鳜胃蛋白酶原基因cdna全长的克隆与序列分析

《鳜胃蛋白酶原基因cdna全长的克隆与序列分析》由会员上传分享，免费在线阅读，更多相关内容在行业资料-天天文库。

1、鳜胃蛋白酶原基因cDNA全长的克隆与序列分析吴雪峰，赵金良（上海水产大学农业部水产种质资源与养殖生态重点开放实验室，上海200090）关键词：鳜；胃蛋白酶原；cDNA末端快速扩增；克隆；序列分析中图分类号：S917文献标识码：ACloningandsequencingofthefull-lengthcDNAofpepsinogengenefromtheMandarinfish，SinipercachuatsiWUXue-Feng,ZHAOJin-Liang(KeyLaboratoryofAquati

2、cGeneticResourcesandAquaculturalEcosystemCertificatedbytheMinistryofAgriculture,ShanghaiFisheriesUniversity,Shanghai200090,China)Abstract:Pepsinogenisonekindofgastricdigestiveproteinasesbelonglingtoafamilyofasparticproteinases,whichissynthesizedinthega

3、stricmucosaofvertebratesandconvertedtopepsinsintheacidicenvironmentofgastricjuice.Themandarinfish(Sinipercachuatsi)isatypicalcarnivorousfishwhichonlypreyonsmalllivefishesafterhatching.Inordertounderstandthemolecularcharactersofpepsinogeninfish,thefull-

4、lengthcDNAofpepsinogengeneofS.chuatsiwasclonedbymeansofRT-PCRandrapidamplificationofcDNAends(RACE)andsequenced.Theresultrevealsa1367bpcDNAfull-lengthsequencecontaining43bp5′-untranslatedregion,187bp3′-untranslatedregionand1137bpopenreadingframe(ORF),wh

5、ichencodes378aminoacidswithcharacteristicsignalpeptidesandactivationpeptides.ThepepsinogenofS.chuatsialsohashighlyconservedaminoacidresiduesessentialforcatalyticactivityandconformationalmaintenance,whicharetwoessentialaspartylresiduesintheactivesiteand

6、sixcysteineresiduesinvolvedinformingthreedisulphidebonds.ThehomologyofaminoacidsequencesbetweenpepsinogenofS.chuatsiandothervertebratepepsinogensrangesfrom59.9%to91.2%,whichindicatesthepepsinogengeneishighlyconservedinevolution.TheNJphylogenetictreeofv

7、ertebratesfromtheaminoacidssequencesofpepsinogenshowedallfishwereclusteredasagroup,thepepsinogenofS.chuatsiwasthemostclosetopepsinogenAformⅡaofwinterflounder.ThesuccessfulcloningofpepsinogengenefromS.chuatsinotonlylaysthefoundationforfurtherstudyofitst

8、emporalandspatialexpression,butalsoprovidesnewmaterialforthemolecularcharacteristicandevolutionoffishpepsinogen.Keywords:Sinipercachuatsi;pepsinogen;rapidamplificationofcDNAends;cloning;sequenceanalysis收稿日期：2007-07-19资助项目：上海市重点学科建设项目(Y1