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1、基因工程外文翻译(中英对照)Retrovirus-mediatedgenetransferandexpressioncloning:PowerfultoolsinfunctionalgenomicsMostofthehumangenomehasnowbeensequencedandabout30,000potentialopenreadingframeshavebeenidentified,indicatingthatweusethese30,000genestofunctionallyorganizeourbiologicactivities.Howeve
2、r,functionsofmanygenesarestillunknowndespiteintensiveeffortsusingbioinformaticsaswellastransgenicandknockoutmice.Retrovirus-mediatedgenetransferisapowerfultoolthatcanbeusedtounderstandgenefunctions.Wehavedevelopedavarietyofretrovirusvectorsandefficientpackagingcelllinesthathavefaci
3、litatedthedevelopmentofefficientfunctionalexpressioncloningmethods.Inthisreview,wedescriberetrovirus-mediatedstrategiesusedforinvestigationofgenefunctionsandfunction-basedscreeningstrategies2003InternationalSocietyforExperimentalHematology.PublishedbyElsevierInc.摘要:人类基因组的大部分现在已经测序完
4、成,大约30,000潜在的开放阅读框已经确定,表明我们使用这30,000个基因管理我们的生物学活和功能性。然而,许多基因的功能仍是未知尽管在生物信息学上以及转基因和基因敲除小鼠做出了大量的努力工作。逆转录病毒介导的基因转移是一个功能强大的工具,可以用来了解基因的功能。我们已经开发出多种逆转录病毒载体和高效的包装细胞系,有利于发展高效功能表达克隆的方法。本文中,我们描述了逆转录病毒介导的方法,用于基因功能和功能为基础的筛选策略的研究。2003InternationalSocietyforExperimentalHematology.PublishedbyEls
5、evierInc.Function-basedgenecloningItwasonly30yearsagothatrecombinantDNAtechnologywasinitiated[1].Theglobingenewasclonedasthefirstmammaliangenein1976thatwasreversetranscribedfromthepurifiedmRNAforglobin[2].ForlessabundantmRNAs,thecDNAswerefrequentlyisolatedbasedontheaminoacidsequenc
6、esofpurifiedproteins.Inlate1970s,amethodcalledhybridselectionwasdeveloped.TheprincipleofthismethodwastodetectproteinstranslatedfromthemRNAhybridizedtoaparticularpoolofsubdividedcDNAlibraryfixedonnitrocellulosemembranes,therebyidentifyingapoolthatcontainsacDNAofinterest.Xenopusoocyt
7、ewasusedforproductionofproteins.Thistypeofexperimentiscalled“expressioncloning,”whichmeans“cloningofcDNAbydetectionofproteinsexpressedfromcDNAlibraries.”ThisstrategyissuitableforisolationofrarecDNAs,suchascDNAsforcytokinesandcytokinereceptors.Levelsoftheproteinexpressionarelow,buta
8、smallamountofproteinisenou
