基因工程第五章（The fifth chapter of gene engineering）.docVIP

下载本文档

2
0
约 11页
2017-08-17 发布于河南
举报
版权申诉

基因工程第五章（The fifth chapter of gene engineering）.doc

1、有哪些信誉好的足球投注网站（book118）网站文档一经付费（服务费），不意味着购买了该文档的版权，仅供个人/单位学习、研究之用，不得用于商业用途，未经授权，严禁复制、发行、汇编、翻译或者网络传播等，侵权必究。。
2、本站所有内容均由合作方或网友上传，本站不对文档的完整性、权威性及其观点立场正确性做任何保证或承诺！文档内容仅供研究参考，付费前请自行鉴别。如您付费，意味着您自己接受本站规则且自行承担风险，本站不退款、不进行额外附加服务；查看《如何避免下载的几个坑》。如果您已付费下载过本站文档，您可以点击这里二次下载。
3、如文档侵犯商业秘密、侵犯著作权、侵犯人身权等，请点击“版权申诉”（推荐），也可以打举报电话：400-050-0827(电话支持时间：9:00-18:30)。
4、该文档为VIP文档，如果想要下载，成为VIP会员后，下载免费。
5、成为VIP后，下载本文档将扣除1次下载权益。下载后，不支持退款、换文档。如有疑问请联系我们。
6、成为VIP后，您将拥有八大权益，权益包括：VIP文档下载权益、阅读免打扰、文档格式转换、高级专利检索、专属身份标志、高级客服、多端互通、版权登记。
7、VIP文档为合作方或网友上传，每下载1次，网站将根据用户上传文档的质量评分、类型等，对文档贡献者给予高额补贴、流量扶持。如果你也想贡献VIP文档。上传文档

基因工程第五章（The fifth chapter of gene engineering）

基因工程第五章（The fifth chapter of gene engineering） In vitro recombination and gene conversion in Chapter fifth The first section is connected DNA segments in vitro Section second of the recombinant was introduced into bacterial cells Connected to a sticky end, (Cohesive end ligation sticky ends connected): With sticky ends of two double stranded DNA molecules in DNA ligase, connected into a hybrid double stranded DNA. Advantage Time saving, convenient operation?; Easy recovery of exogenous fragment?; The problem Carriers ring? The insert can be inserted into the two? 2. different restriction enzyme connection at the end of the stick ? molecular carrier and gene has two identical restriction sites The vector and the target gene? Only one the same restriction enzyme recognition site No? The same restriction enzyme recognition sites, have isocaudarner The Sal fragment (CAGCTG) Xho fragment (GAGCTC) If you are not above?? At the end of two, flat connection Flat end joining (blunt end ligation): In the T4DNA ligase, the two has a flat end of the double stranded DNA molecules connected into DNA hybrid molecules. Advantage You can use any T4 ligase DNA flat end 5? - prominent sticky ends up The available Klenow polymerase to complete 3? - prominent sticky ends cut Available T4 DNA polymerase Xiaoping The main problem The connection efficiency is low ? high concentrations of substrate and T4 DNA ligase Adding coagulant: such as? It is not easy to recover the exogenous DNA fragment Two-way insertion Three, the modified sticky end connections Homopolymeric tail method?: Homopolymeric tail (homopolymer tails joining) is connected by terminal transferase in carrier and exogenous double stranded DNA 3ends with a oligonucleotid, made of artificial viscosity and then in the end, DNA ligase, connected into recombinant DNA. Advantage The effect of self loop does not occur; The higher connection efficiency; The problems Complex operation? ? gene difficult recovery May affect gene express