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Advances in Pig Genomics and Functional Gene Discovery

Comparative and Functional Genomics

Comp Funct Genom2003;4:266–270.

Published online1April2003in Wiley InterScience(https://www.sodocs.net/doc/0d865320.html,).DOI:10.1002/cfg.261 Conference Review

Advances in pig genomics and functional gene discovery

Max F.Rothschild*

Department of Animal Science,Center for Integrated Animal Genomics,Iowa State University Ames,IA50011,USA

*Correspondence to:

Max F.Rothschild,2255Kildee Hall,Department of Animal Science,Iowa State University, Ames,IA50011,USA.

E-mail:mfrothsc@https://www.sodocs.net/doc/0d865320.html,

Received:28January2003 Accepted:28January2003Abstract

Advances in pig gene identi?cation,mapping and functional analysis have continued to make rapid progress.The porcine genetic linkage map now has nearly3000loci, including several hundred genes,and is likely to expand considerably in the next few years,with many more genes and ampli?ed fragment length polymorphism(AFLP) markers being added to the map.The physical genetic map is also growing rapidly and has over3000genes and markers.Several recent quantitative trait loci(QTL) scans and candidate gene analyses have identi?ed important chromosomal regions and individual genes associated with traits of economic interest.The commercial pig industry is actively using this information and traditional performance information to improve pig production by marker-assisted selection(MAS).Research to study the co-expression of thousands of genes is now advancing and methods to combine these approaches to aid in gene discovery are under way.The pig’s role in xenotransplantation and biomedical research makes the study of its genome important for the study of human disease.This review will brie?y describe advances made, directions for future research and the implications for both the pig industry and human health.Copyright 2003John Wiley&Sons,Ltd.

Keywords:pig;gene identi?cation;economic traits;functional genomics

Introduction

The pig was among one of the?rst animals domes-ticated over7000years ago and pork is the major red meat consumed(43%)worldwide[23].Further-more,the pig has served as an important model system for human health and represents a signif-icant future source of organs for transplantation. Efforts to unravel the pig genome began in the early1990s with the development of the PiGMaP gene mapping project[1],which was initiated in Europe and was funded by the European Eco-nomic Community.PiGMaP involved18Euro-pean labs and a total of7other labs from the USA,Japan and Australia.In the USA,the USDA launched two efforts.First,the USDA–ARS(US Department of Agriculture–Agricultural Research Service)began a sizeable gene mapping project [21]at the Meat Animal Research Center in Clay Center,Nebraska.Second,the National Animal Genome Research Program was developed under the direction of USDA–CSREES(Cooperative State Research Education and Extension Service) in1993.This program was designed to provide a structure that included genome coordinators that would stimulate facilitation and collaboration of gene mapping in all species,including pigs.Scien-tists from state and private universities and federal labs cooperatively created a Swine Genome Tech-nical Committee,which has met in recent years at the Plant and Animal Genome(PAG)Meetings. The US Pig Genome Coordinator activities,in con-cert with activities of the USDA–ARS and inter-national gene mapping projects,such as PiGMaP and others,have allowed the status of the pig gene map to evolve more quickly and developments in functional genomics to advance rapidly in the last several years.

Pig genomics and functional gene discovery267

Gene mapping

New gene markers consisting of microsatellites, ampli?ed fragment length polymorphisms(AFLPs) and single nucleotide polymorphisms(SNPs)con-tinue to be identi?ed and mapped and some inte-gration of the maps continues to have taken place as quantitative trait maps are expanded.The largest single map contains about1200markers[21]but no new large-scale maps have been published recently. In total there are over924genes and1641mark-ers in the database(https://www.sodocs.net/doc/0d865320.html,/browser? species=pig).There is a developing AFLP map with about3000AFLPs that is likely to be added to the PiGMaP linkage map some time in the future. Integration of the linkage,cytogenetic and physical maps is well under way with the development and use of chromosome painting[14],a somatic cell hybrid map[28]and a7000rad radiation hybrid (RH)panel(ImpRH)[30,15].This RH map now contains nearly3000markers including microsatel-lites,and over2000new expressed sequence tags (ESTs),of which many are human orthologues and enable comparative mapping[20,24].Contin-ued use of these resources and development of an advanced12000rad RH map are under way [29].This will aid the rapidly developing com-parative map,which will accelerate the identi?ca-tion of the genes explaining variation in traits of interest,either those identi?ed by QTL studies or through direct approaches,such as gene association analyses.

Database activities

Signi?cant pig bioinformatics efforts have been initiated by the Roslin Institute,Scotland(www. https://www.sodocs.net/doc/0d865320.html,)and to a lesser extent in the USA(https://www.sodocs.net/doc/0d865320.html,)to support the pig genome efforts and display the gene maps[2].PiG-BASE,which can be reached through these sites, has several features,including pig gene mapping references with over1093citations in the database and gene maps with https://www.sodocs.net/doc/0d865320.html,st year there were over2million hits at these pig genome sites.Additional websites exist for the cytoge-netic map of the pig(http://www.toulouse.inra.fr/ lgc/pig/cyto/cyto.htm)and the RH panel map (http://www.toulouse.inra.fr/lgc/pig/RH/Menu-chr.htm).A comparative map is also on the web (http://www.toulouse.inra.fr/lgc/pig/compare/ compare.htm).In addition,a new EST database (https://www.sodocs.net/doc/0d865320.html,)has been devel-oped and should become a similarly useful resource. It is now accessible and contains over98988pig EST entries and further development will continue. Other useful gene tools are available from the US pig genome website(http://www.genome.iastate. edu).

QTL and candidate genes

Pork production requires ef?cient growth rate, reduced feed intake,carcass merit,meat quality and high levels of reproductive success and https://www.sodocs.net/doc/0d865320.html,ing both commercial and exotic pig breeds, researchers have initiated experiments to iden-tify quantitative trait loci(QTLs)affecting these traits.A large number of QTLs have been reported on nearly all chromosomes for growth,carcass and meat quality traits and several chromosomes for reproduction[3].The QTLs affecting immune response traits and disease resistance are far less numerous.This is an area where gene expression approaches may be particularly valuable.Follow-ing discoveries of imprinted genes in other species, researchers have expanded their projects to?nd imprinted and origin-of-parent effects[10].In par-ticular,one such region on chromosome2has been intensively investigated[12]and IGF2impli-cated in causing a major effect in muscle mass. The researchers cleverly employed a haplotype-sharing strategy analysis combined with marker-assisted segregation analysis to position the QTL within a500kb region.The causal quantitative trait nucleotide(QTN)was revealed after investigating over180SNPs and this work clearly points to the need for careful analysis of all gene regions and the proper animals and phenotypic information.Further evidence for imprinted regions and genes are likely to be found now that these approaches have been developed.

Candidate genes analyses have been employed to investigate a variety of traits.To date,signi?-cant associations have been demonstrated for can-didate genes for litter size(ESR,PRLR,RBP4), growth(MC4R),meat quality(PRKAG3),disease resistance(FUT1,SLA,NRAMP)and coat color (KIT,MC1R)[3].The commercial pig industry is actively using this gene marker information in

268M.F.Rothschild

combination with traditional performance informa-tion to improve pig production by marker-assisted selection.Positional candidate gene analysis con-tinues to be used to elucidate other known QTLs and has recently been useful in uncovering QTN mutations in PRKAG3that affect pH and drip loss [6]and in CAST that affect tenderness[7].It is likely that,as QTL experiments are expanded,addi-tional positional candidates will be identi?ed and the causative QTN discovered.

Sequencing efforts

Research to date suggests that the porcine genome has a similar chromosomal organization(2n=38, including meta-and acrocentric chromosomes), size(3×109bp),and complexity to the human genome.As with other species,researchers have generated ESTs from cDNA clones randomly picked from libraries from many tissues.These projects have varied in size and in the tissues used[8,17,19,26,27].The largest of these types of projects published to date was sponsored by the USDA and reported the sequencing and initial analysis of66245ESTs[11].In addition,21499 sequences from reproductive tissue were produced by a consortium of several research groups[24].At present,there are approximately120000sequences in GenBank,and in the October2002TIGR release there were17350clusters and31000singletons. More deposits of5000–10000EST sequences are expected soon.Most importantly,however, a major Sino-Danish effort to sequence the pig genome(http://www.piggenome.dk/)has resulted in approximately700000EST sequences that are expected to be deposited in the database in the next6–8months.The data obtained by sequencing these large numbers of ESTs will continue to help assist comparative mapping efforts,candidate gene discovery and expression analysis.

Following the request of the NIH,a number of species have submitted requests to be considered for sequencing efforts.A‘White Paper’[22]was submitted to NHGRI recently that outlined the role the pig plays agriculturally,as well as a model for human biology.In addition to the efforts of the authors,the White Paper received solid backing from colleagues from several countries and from industry personnel from many companies and orga-nizations.A cooperative project to develop a BAC map using the existing BAC library resources with approximately35×coverage[22]has progressed nicely.It appears that the pig genome sequenc-ing effort will receive a‘high priority ranking’but, despite these efforts,suf?cient funding remains in question.

Functional analysis

To better understand the physiological complex-ity of the pig transcriptome,expression and/or functional gene analysis needs to be undertaken. Initially such research was done using a lim-ited number of genes and techniques,such as Northern analysis and differential display PCR [13,25].Other approaches have included quanti-tative real-time PCR to determine mRNA levels for immune response and disease infection lev-els[9,18].These approaches,while quite useful, have proved to be limited in the numbers of genes that can be considered.Other approaches have included use of limited numbers of cDNAs on macroarrays[31].Given the initial lack of devel-opment of large-scale cDNA arrays for the pig, human arrays have been tested and used[13,16]. Experiments with such materials have proved ini-tially valuable,as reproducibility was generally high and results were reasonable.However,the recent advent of large numbers of pig ESTs has allowed for large-scale expression analysis using porcine materials only.Pomp and colleagues[4,5] have used cDNA derived from ovary and follicu-lar RNA from animals from either an index line selected for higher litter size or a control line, and co-hybridized them with4600follicle-derived probes to study gene expression patterns related to reproductive ef?ciency.Other projects exist includ-ing two large-scale efforts in Europe.The?rst European Community-supported project is called PathoCHIP(https://www.sodocs.net/doc/0d865320.html,) and uses spotted cDNA arrays for disease organ-ism and immune response genes,while the second, called QualityPorkGENES(https://www.sodocs.net/doc/0d865320.html,)looks at the co-expression of genes related to meat quality.Cooperative efforts by the US Pig Genome Coordinator and US and International researchers have now been directed to developing a?rst stage cDNA or oligo spotted array for the pig genome and human biomedical community.It is expected that such an array will be commercially

Pig genomics and functional gene discovery269

available in mid-2003.This array and others to be developed later will advance functional analysis in the pig.

Conclusions

Understanding the complexity of the pig genome for both agricultural purposes and for its impor-tance to human biomedical concerns remains a signi?cant challenge.In the past decade,large-scale gene and trait identi?cation and mapping have taken place and a number of gene tests to improve pork production are in use in the pig industry. Sequencing and expression analysis have been ini-tiated and offer new avenues to understand the biological complexity of the pig.No longer does the pig genomics community rely solely on devel-opments from other organisms,such as the human and the mouse.This sentiment was also shared by Nobel laureate Dr Sydney Brenner at this year’s PAG meeting,where Dr Brenner stated,‘more seri-ous people work on important things’,expressing his belief that scientists should now focus on their primary species and not model systems.The state of the art of pig gene discovery and functional genomics clearly demonstrates such commitment and progress.

Acknowledgements

This is a journal paper of the Iowa Agriculture and Home Economics Experiment Station,Ames,IA,USA, Project No.3148,and was supported in part by Hatch Act and State of Iowa funds and funding from NRSP-8and the USDA/CSREES Pig Genome Coordination program. The efforts of all scientists involved in the pig gene mapping and genomics programs worldwide are gratefully acknowledged,including efforts by Alan Archibald,Andy Law and colleagues,Roslin Institute,and Zhiliang Hu to support database https://www.sodocs.net/doc/0d865320.html,ments and information supplied by H.Cheng,C.Ernst,M.Georges,G.Plastow, D.Pomp and C.Tuggle are appreciated. References

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peppa_pig 第一季英文文本

1. Muddy puddles This is my little brother, George. This is Mummy Pig. And this is Daddy Pig. Peppa Pig. Muddy puddles. It is raining today. So, Peppa and George cannot play outside. Daddy, it's stopped raining. Can we go out to play? Al right, run along you two. Peppa loves jumping in muddy puddles. I love muddy puddles. Peppa, if you jump in muddy puddles, you must wear your boots. Sorry, Mummy. George likes to jump in muddy puddles, too. George. If you jump in muddy puddles, you must wear your boots. Peppa likes to look after her little brother, George. George, let's find some more puddles. Peppa and George are having a lot of fun. Peppa has found a little puddle. George has found a big puddle. Look, George. There's a really big puddle. George wants to jump into the big puddle first. Stop, George. I must check if it's safe for you. Good. It is safe for you. Sorry, George. It's only mud. Peppa and George love jumping in muddy puddles. Come on, George. Let's go and show Daddy.

小猪佩奇第一季第一家中英文字幕

1.Muddy Puddles 泥水坑 It is raining today. So, Peppa and George cannot play outside. 今天下雨了，所以佩奇和乔治不能去外面玩耍。 Daddy,it's stopped raining. 爸爸，雨停了。 Can we go out to play? 我们可以出去玩了吗？ Alright, run along you two. 好的，你们两个一起去吧。 Peppa loves jumping in muddy puddles. 佩奇喜欢在泥坑里玩。 I I love muddy puddles. 我喜欢在泥坑里玩。 Peppa. If you jump in muddy puddles, you must wear your boots. 如果你在泥泞的水坑里跳，你必须穿上你的靴子。 Sorry, Mummy. 对不起，妈咪。 George likes to jump in muddy puddles, too. 乔治也喜欢跳在泥泞的水坑。 George. If you jump in muddy puddles, you must wear your boots. 乔治。如果你在泥泞的水坑里跳，你必须穿上你的靴子。 Peppa likes to look after her little brother, George. 佩奇喜欢照顾她的弟弟，乔治。 George, let's find some more puddles. 乔治，让我们找到一些更多的水坑。 Peppa and George are having a lot of fun. 佩奇和乔治玩得很开心。 Peppa has found a little puddle. George has found a big puddle. 佩奇发现一个小水坑。乔治发现了一个大水坑。 Look, George. There's a really big puddle. 你看，乔治。有一个真正的大水坑。 George wants to jump into the big puddle first. 乔治希望第一个跳大水坑。 Stop, George. I must check if it's safe for you. Good. It is safe for you. 停止，乔治。我必须检查一下这个对你来说是不是安全的。很好。它很安全。Sorry, George. It's only mud. 对不起，乔治。那只是泥而已。 Peppa and George love jumping in muddy puddles. 佩奇和乔治很爱在泥泞的水坑里跳。 Come on, George. Let's go and show Daddy! Goodness me.

综合实践活动课程介绍

综合实践活动是普通高中新课程方案中规定的一门必修课程。它包括研究性学习、社区服务、社会实践三个部分，总共配置 23 个学分，其中研究性学习占 15 个学分，社区服务和社会实践分别为 2 个和 6 个学分。研究性学习是普通高中阶段综合实践活动课程的最重要的内容。作为指导教师，要充分认识到研究性学习对学生的发展价值和意义，把握研究性学习的基本规范，掌握研究性学习的各种操作方法，提高指导的有效性。综合实践活动课程是教师引导下，学生自主进行的综合性学习活动，是基于学生的经验，密切联系学生自身生活和社会实际，体现对知识的综合应用的实践性课程。研究性学习是学生在教师指导下，从自然、社会和学生自身生活中选择和确定研究专题，主动获取知识、应用知识、解决问题的学习活动。一、综合实践活动的课程性质【课程解读】作为普通高中的一门必修课程，综合实践活动课程超越具有严密的知识体系和技能体系的学科界限，它不是一门学科课程，而是有着不同于学科课程的规定性的课程。一般来说，综合实践活动课程具有如下基本规定。 1. 综合实践活动课程是一种经验性课程。所谓经验性课程，是指一方面课程的设计是以学生的生活经验、生活背景、生活中感受到的问题与需要为中心的，强调学生经验在课程中的独特地位和价值；另一方面，课程的实施是建立在学生的活动经验和活动过程基础之上的，强调学生的亲身经历和参与。经验课程的基本规定之一是强调学生经验的发展价值。一般来说，经验课程反对把书本知识作为现存的结论或定论直接告诉学生，而是通过经验获得、经验的重新组合或经验重塑，使学生获得成长的意义。经验课程基本组织方式是“问题中心”，注重学生通过自主的问题探究与问题解决实现课程的发展价值。可以说，在教育史上，经验课程从来都是与学科本位或知识本位课程相冲突的。经验课程重视的不是系统的知识的获得，而是经验重组与改造过程中学生兴趣、爱好、能力、情感、价值观等方面的发展，以及在经验生成与改造过程中学生的批判性思维的发展乃至生活方式的变革。经验课程的基本出发点是学生在教育中的主体地位。应该说，历史上的各种经验课程的共同之处就在于强调学生在教育中的主体地位和自主发展的能力。学生是经验者、是经验改造者，是自主发展者。经验课程设计的意义在于为学生的自主发展提供一种以问题为中心的发展环境。如果在经验课程中学生的主体地位得不到尊重和发挥，那么，其课程价值就难以得到实现。经验课程的设计面对的不是符号系统，而是鲜活的生活世界，面向学生的生活情景，面向学生的生活经历和生活领域，并以整合的方式组织经验，是经验课程的基本特点之一。在生活情景中发现问题、明确问题，通过自主探究和实践，形成问题解决的能力和组织生存的能力。日本新课程中的“综合学习时间”的基本活动领域之一就是自主生存。因为学生的发展不应该完全是在预设的课堂教学和学科知识的传递过程中实现的，如何面向现实的生活，引导学生通过自主实践、批判反思来获得发展，成为经验课程关注的核心问题。

综合实践主题报告

长兴县2016年中小学生综合实践活动学习成果展示评奖活动推荐表

[内容摘要] 根雕作为一种民间艺术，在中国的发展历史源远流长，博大精深。在现代社会中，它以其独具匠心、妙趣天成的艺术感染力，受到越来越多人的喜爱。在我们长兴也有许多民间艺人和根雕爱好者，默默传承着根雕这一传统文化，水口的东方梅园也在筹备根雕博物馆。为了让长兴的孩子更多地了解根雕，学会基本制作根雕的方法，我们开展了“走进根雕世界，聆听文化回音”综合实践活动。在本次活动从寒假一直延续到今年3月份，过程漫长，活动丰富多彩。活动初期，同学们广泛调查与根雕相关的资料，拜访了根雕艺人，参观了根雕博物馆。活动中期在开展了一系列探究、考察、采掘、制作、拓展、创新的综合实践活动，活动后期进行了反思和成果展示。在整个活动的过程中，同学们体验了寻根、挖根、制作根雕的过程，还对根雕作品进行了改造和创新。不仅促进学生亲近自然，保护环境，而且在合作、探究的过程的能力，提高了我们创新意识和动手能力，将传统文化与实践创新紧密结合是我们这次活动的最大收获。

“走进根雕世界，聆听文化回音”主题活动报告一、活动主题的产生 1．综合实践活动要因地制宜，因时制宜，并且具有一定的活动意义。根雕作为一种传统文化，为世人所喜爱。为了传承根雕艺术，认识根雕的类型，学习根雕的制作工艺，拓展我们实践和创新的能力，我们实小303的孩子们以根雕为载体，开展综合实践活动。 2．综合实践活动要以学生为主体，符合学生的兴趣爱好。从我们综合实践活动前的调查表显示出：同学们对根雕很感兴趣。而且根雕的创新过程中也会产生很多有意义的作品，开展有趣的活动，这些都将吸引孩子们将实践进行下去。 3．根雕的实践活动中有很多操作的环节可以锻炼学生探究的能力和动手实践的能力。还促进学生在合作、探究的过程中增强信息搜索和整理的能力。在改进和创新的过程中，提高了我们创新思维和动手能力。 4.根雕展示的活动可以让社会上的人们增加对根雕的认识和了解，起到传播传统文化的作用，培养学生制作传统工艺的兴趣。二、主题活动的具体目标知识目标： 1.了解根雕的文化背景和选材需要。 2.了解根雕的制作工艺和选材要求。能力目标： 1. 培养学生调查访问、收集处理信息、语言表达的能力以及团队合作的精神。 2.学会根雕的制作过程，能制作简单的根雕艺术品。 3.通过活动培养学生的兴趣爱好，陶冶情操。并能对根雕制作进行改进和创新。情感态度与价值观目标： 1.培养学生亲近自然，热爱传统文化的人生态度。 2.培养学生勇于创新、乐于学习的态度。 3.激发探究的热情，培养探究能力。三、活动实施的具体过程：第一阶段发动倡议阶段活动时间：1周一、活动的前期准备 1．讨论实践的课题。

peppa pig第一季 41 shopping 字幕翻译

peppa pig第一季 41 shopping 字幕翻译Shopping 旁白:Peppa and George are going shopping! Peppa and George like shopping. George loves sitting in the trolley. So does Peppa. 旁白:佩佩和乔治要去买东西～佩佩和乔治喜欢买东西。乔治喜欢坐在推车里，佩佩也一样 Peppa: Daddy, can I sit in the trolley ,too? 佩佩:我也想坐在推车里。Daddy: You're too big for the trolley, Peppa. 爸爸:佩佩，你太大了不能坐推车了。 Peppa: oh! 佩佩:哦.. Daddy:But you can help with the shopping. 爸爸:不过你可以帮忙找东西Peppa:oh!goody! 佩佩:好棒哦 Mother:We're got four things on the list. 妈妈:我们的购物单上有四样东西 . Daddy:Tomatoes, spaghetti, onions and fruit. 爸爸:番茄、意大利面、洋葱和水果 Peppa:I'll find it all! This way! 佩佩:我都找得到，走这边。旁白:Peppa and George love shopping. 旁白:佩佩和乔治最喜欢来买东西了。Mother: Peppa first,we need tomatoes. 妈妈:佩佩第一样东西是番茄 Peppa: Hmmm! I can see them! I can see them! Here are the tomatoes, Mummy. 佩佩:哦...我看到了，我看到番茄了。番茄在这里妈咪。 Mummy:Well done,Pepaa! 妈妈:你好厉害啊。

Peppa Pig

Peppa Pig第1集：吹泡泡【学习笔记】重点词组讲解 1. Bubble泡泡吹泡泡：blow bubbles 例句：Peppa is blowing bubbles in her drink. 佩佩正往她的饮料里吹泡泡。 2. Orange Juice 橘子汁柠檬汽水：lemonade 3. too 也…… 例句：George wants to blow bubbles too. 乔治也想吹泡泡。 4. play with your food 玩你的食物例句：Geroge, you really shoudn't play with your food. 乔治，你真的不应该玩你的食物。这句话经常可以听到外国的父母对自己的孩子说哦！ 5. bubble mixture泡泡液 6. take a deep breath 深吸一口气 7. Now it's your turn. 现在该你了！ 8. running out of breath 呼吸不过来了、喘不过气来了 9. be used up 用完了例句：The bubble mixture is all used up. 泡泡液都用完了。 10. tennis racket网球拍 11. muddy puddle泥坑

Peppa Pig第2集：泰迪的一天【小Car笔记】重点词组讲解 1, stuff 物品、东西例句：have you go your stuff ready? 你们都带好自己的东西了吗？ 2, look after 照看、看管例句：You must look after them. 你们必须好好照看他们。 3, leave……（behind）忘带、遗留例句：George left Mr. Dinosaur behind. 乔治忘记带恐龙先生了。 4, Here we go again! 我们又出发了！ 5, day off 放假、假日例句：It's my day off. 今天我放假。 6, picnic lunch 野餐例句：We have a picnic lunch. 我们来野餐。 7, leave the table 离席例句：Please may George and I leave the table? 我和乔治能离席去玩吗？8, play with 与 ... 一起玩例句：I play with Zebi. 我要和泽比一起玩。 9, I got the picnic toy sets. 我带了一套野餐玩具。 10, pretend假装、装作例句：We got pretend cake. 我们假装着是蛋糕。 11, The best cake ever! 有史以来最好吃的蛋糕！ 12, Home time。回家的时间到了/该回家了。 13, Oh, no. You shouldn't be here Teddy! 噢，不。你不应该在这的，泰迪。

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小猪佩奇第一季1-5集英文字母

Peppa Pig 第一季1 Muddy puddles This is my little brother, George. This is Mummy Pig. And this is Daddy Pig. Peppa pig. Muddy Puddles. It is raining today. So, Peppa and George cannot play outside. Daddy, it's stopped raining. Can we go out to play? Al right, run along you two. Peppa loves jumping in muddy puddles. I love muddy puddles. Peppa. If you jump in muddy puddles, you must wear your boots. Sorry, Mummy. George likes to jump in muddy puddles, too. George. If you jump in muddy puddles, you must wear your boots. Peppa likes to look after her little brother, George. George, let's find some more puddles. Peppa and George are having a lot of fun. Peppa has found a little puddle. George has found a big puddle. Look, George. There's a really big puddle. George wants to jump into the big puddle first. Stop, George. I must check if it's safe for you. Good. It is safe for you. Sorry, George. It's only mud. Peppa and George love jumping in muddy puddles. Come on, George. Let's go and show Daddy. Goodness me. Daddy. Daddy. Guess what we've been doing. Let me think... Have you been watching television? No. No. Daddy. Have you just had a bath? No. No. I know. You've been jumping in muddy puddles. Yes. Yes. Daddy. We've been jumping in muddy puddles. Ho. Ho. And look at the mess you're in. Oooh... Oh, well, it's only mud. Let's clean up quickly before Mummy sees the mess. Daddy, when we've cleaned up, will you and Mummy Come and play, too? Yes, we can all play in the garden. Peppa and George are wearing their boots. Mummy and Daddy are wearing their boots. Peppa loves jumping up and down in muddy puddles. Everyone loves jumping up and down in muddy puddles. Oh, Daddy pig, look at the mess you're in. It's only mud. I'm Peppa Pig. This is my little brother, George. This is Mummy Pig. 2 Mr. Dinosaur is Lost. George's favourite toy is Mr. Dinosaur. DineDine SawSaw. George loves Mr. Dinosaur. Grrrrrrrrrrrrrrrr.. Sometimes, George Iikes to scare Peppa with Mr. Dinosaur. Grrrrrrrrrrrrrrrrrrrr. Eeek. Too scary. At suppertime, Mr. Dinosaur sits next to George. I beg your pardon. Was that you George, or was it Mr. Dinosaur? DineDine SawSaw. At bath time, George shares his bath with Mr. Dinosaur.

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分子生物学基础和技术教学大纲(精)

分子生物学基础和技术教学大纲（适用于医学检验和医学相关专业）课程性质与目的分子生物学是医学领域发展最快的学科之一，日新月异的技术使它逐渐成为医学发展的重要支柱。随着本世纪初人类基因组计划的完成，医学发展进入了一个全新的时代。疾病基因的不断发现和克隆，使人们对疾病的认识也不断深入，而这些重大的医学进步离不开技术上的更新和发展，生物芯片技术、基因测序技术、毛细管电泳技术等，每一次技术的进步都为分子生物学的发展提供了有力的保障。分子生物学技术是一门重要的基础和应用课程，教学方式目前主要以理论课程为主，分基础理论和基础技术两个部分，重点讲述分子生物学检验技术的基础理论和基础知识，并引入近年发展的新理论、新技术，使学生了解和学习最新进展和相关内容。同时分子生物学技术最主要的作用是作为研究医学的一种媒介和工具，具有很强的实践性，其基本知识和理论来源于科学实验，因此现在现针对本科学生开展了分子生物学实验课程，实验教学是强化理论课的重要方式，是培养医学生实验科学概念和实验技能的重要途径，通过综合性的实验可以强化学生对理论的深入理解和实际运用，可以更全面直观的分析理论知识。更重要的是，实验教学是培养学生综合分析和解决问题的能力以及科学创新能力的重要方式。本课程的目的是通过分子生物学重要技术的学习，使学生掌握一门可运用于医学研究的技术和工具，了解医学发展的最新进展和前沿技术，通过理论与实践的结合将分子生物学融入医学研究的各方面，分析疾病基因、从分子水平分析疾病发生的原因、跟踪疾病发展过程、检测感染人类的病原生物以及未来根据个体化治疗奠定理论和技术基础。课程的设置与要求本课程是在学生系统学习了前期课程的基础上由检验系临床化学教研室负责开设的，与本课程相关的基础课程有生物化学和生化技术等。本课程分为理论课程和实验课程两部分。理论课主要包括基础理论和基本技术，基础理论主要讲授基因和基因组、原核生物和真核生物基因组、人类基因组计划、蛋白质组学、肿瘤分子生物学等；基本技术包括了核酸提取、DNA重组技术、核酸干扰技术、核酸分子杂交、聚合酶链反应、DNA芯片等。实验操作可以使学生将晦涩的理论知识更加融会贯通，因此针对本科学生选择性开设了实验课程——基

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粉红小猪妹Peppa_Pig第二季_01_Bubbles Peppa and George are drinking orange juice. What a noise! Little piggies! "George, look at all the tiny bubbles." I can make bigger bubbles. Peppa is blowing bubbles in her drink. George wants to blow bubbles too. "Peppa, George, you really" shouldn't play with your food! "Sorry, Daddy." "Peppa, if you like bubbles," Why don't you play with your bubble mixture? Yes! My bubble mixture! "George, let's blow bubbles in the garden." Peppa is showing George how to blow bubbles. "First, dip the stick in the mixture." "Then, take a big breath and blow." Heeee... bubble! "George, now it's your turn!" "Dip the stick in the mixture," "hold it up," "take big breath," and blow! "George is trying really hard," but he. S running out of breath! "George, you didn't blow hard enough!" I know a way George can make bubbles. "Dip the stick in the mixture," then lift it up and wave it around. Bubble! It's even better if you run along. Wait for me! "George is having fun making bubbles," and Peppa is having fun popping. I love bubbles! "Oh, dear! The bubble mixture is all used up." "Daddy, Mummy!" There's no more bubble mixture! "Don't worry, Peppa. I have an idea." "Mummy Pig, bring me a bucket, please!" Yes! Daddy Pig. "Peppa, bring me some soap!" "Yes, Daddy Pig." "George, bring me my old tennis racket." "A bucket, some soap, a tennis racket!" What is Daddy Pig doing? "First, I need some water." "Then, the soap." "And now, the tennis racket, please!" Thank you! Now watch this! Woo... Wow! Daddy Pig has made his own bubble mixture. "Come on, everyone!" Be careful! Daddy Pig! Don't slip and fall. I'll be fine! Wow... "Are you Ok, Daddy Pig?" "Yes, I'm fine!" Wow! Daddy Pig has made the biggest bubble ever! Oh dear! I've spilled all my bubble mixture. "Now, we can't make any more bubbles!" But we can play jumping in muddy puddles! Look! Daddy Pig's bubble mixture has made a big muddy puddle! Hurray! Peppa loves jumping up and down in muddy puddles! Look! The bubble mixture has made the puddle all bubbly! Peppa loves jumping up and down in muddy bubbly puddles! Everyone loves jumping up and down in muddy bubbly puddles! 粉红小猪妹Peppa_Pig第二季_02_Teddy_s_day_out Teddy's day out Peppa and her family are going out forthe day. Daddy Pig has made a picnic. "Peppa, George, have you got yourstuff ready?" "Yes, I'm taking teddy!"

PeppaPig小猪佩奇季全英字幕

Peppa Pig第一季 1 Muddy pud dles This is my little brother, George. This is Mummy Pig. And this is Daddy Pig. Peppa pig. Muddy Puddles. It is raining today. So, Peppa and George cannot play outside. Daddy, it's stopped raining. Can we go out to play? Al right, run along you two. Peppa loves jumping in muddy puddles. I love muddy puddles. Peppa. If you jump in muddy puddles, you must wear your boots. Sorry, Mummy. George likes to jump in muddy puddles, too. George. If you jump in muddy puddles, you must wear your boots. Peppa likes to look after her little brother, George. George, let's find some more puddles. Peppa and George are having a lot of fun. Peppa has found a little puddle. George has found a big puddle. Look, George. There's a really big puddle. George wants to jump into the big puddle first. Stop, George. I must check if it's safe for you. Good. It is safe for you. Sorry, George. It's only mud. Peppa and George love jumping in muddy puddles. Come on, George. Let's go and show Daddy. Goodness me. Daddy. Daddy. Guess what we've been doing. Let me think... Have you been watching television? No. No. Daddy. Have you just had a bath? No. No. I know. You've been jumping in muddy puddles. Yes. Yes. Daddy. We've been jumping in muddy puddles. Ho. Ho. And look at the mess you're in. Oooh... Oh, well, it's only mud. Let's clean up quickly before Mummy sees the mess. Daddy, when we've cleaned up, will you and Mummy Come and play, too? Yes, we can all play in the garden. Peppa and George are wearing their boots. Mummy and Daddy are wearing their boots. Peppa loves jumping up and down in muddy puddles.

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药学分子生物学常用分子生物学技术的教学体会药学分子生物学已成为21世纪药学专业高等教育的核心课程，针对药学分子生物学实践性很强的特点，我们对常用分子生物学技术授课内容进行改革，结合教学实际，设置具有特色的理论和实验结合的课程体系。根据此章节授课内容的自身特点和教学中的实际经验，总结四点教学体会：介绍本学科最新前沿动态，提高学生兴趣；注重融会贯通、重点突出，便于学生掌握重难点；应用多媒体教学，对学生进行启发性教学；将自身科研经验和实验实际操作运用到教学中。充分调动学生的学习积极性，提高理论和实验教学的质量。标签：药学分子生物学；生物学技术；课堂教学；教学方法药学分子生物学是在药学、遗传学和生物化学等学科的基础上发展融合形成的新学科；它是将分子生物学的新理论、新技术渗入药学研究领域，从而使药物学研究由化学、药学的培养模式转化成为生命科学、药学和化学相结合的新药模式；同时它还是当代生物科学发展的引擎，是现代生物技术的基石。[1]近年来随着分子生物技术的发展，它的应用领域也在不断拓宽，它与医学和药学方面的交叉也越来越多，因此，分子生物学在今后已经不再只是生物技术专业的必修课，它也成为药学院学生的重要必修的基础课之一。分子生物学主要是从分子水平上阐述生命现象和本质的科学，是现代生命科学的“共同语言”。分子生物学技术把研究技术提高到了基因分子水平，可应用于遗传性疾病的研究和病原体的检测及肿瘤的病因学、发病学、诊断和治疗，新药开发等方面的研究。所以，常用分子生物学技术是现代分子生物学研究的重要核心内容之一。掌握了常用分子生物学技术，并能将理论和实际操作结合，也就相当于掌握了一把从微观世界揭示生物学奥秘的钥匙。经过多年教学，笔者将对此章节的教学体会总结如下。一、介绍本学科最新前沿动态，提高学生兴趣分子生物学是一门发展快速的前沿学科，由其发展带来的成果和研究进展日新月异。由于教材跟不上分子生物学发展速度，在授课时我们及时将最新的分子生物学进展补充到教学内容之中。以基因敲除技术为例，教材主要介绍传统第一代同源重组方法，这种方法是经典基因敲除方法，但效率低（1 per 106 cells），实验周期长，可以说基本被淘汰的方法。随后又出现了锌指核酸酶（ZFN）[2]、TALEN、CRISPR/Cas9等方法。尤其是2012年出现最新CRISPR/Cas9方法，以能够实现任意敲除、成功率高、打靶效率很高、脱靶率高、周期非常快等优点著称。这种方法构建的基因突变动物具有显著高于传统方法的生殖系转移能力，是一种高效、快速、可靠的构建敲除动物模型的新方法，所以在动物模型构建的应用前景将非常广阔。将这些最新的分子生物学科学进展补充到教学内容之中，一方面可以提高学生学习兴趣，另一方面使学生了解本学科最近发展动态，从动态中学习，而非死

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