鳜胃蛋白酶原基因cdna全长的克隆与序列分析

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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(KeyLaboratoryofAquaticGeneticResourcesandAquaculturalEc

2、osystemCertificatedbytheMinistryofAgriculture,ShanghaiFisheriesUniversity,Shanghai200090,China)Abstract:Pepsinogenisonekindofgastricdigestiveproteinasesbelonglingtoafamilyofasparticproteinases,whichissynthesizedinthegastricmucosaofvertebratesandconvertedtopepsinsintheacidicenvironmento

3、fgastricjuice.Themandarinfish(Sinipercachuatsi)isatypicalcarnivorousfishwhichonlypreyonsmalllivefishesafterhatching.Inordertounderstandthemolecularcharactersofpepsinogeninfish,thefull-lengthcDNAofpepsinogengeneofS.chuatsiwasclonedbymeansofRT-PCRandrapidamplificationofcDNAends(RACE)ands

4、equenced.Theresultrevealsa1367bpcDNAfull-lengthsequencecontaining43bp5′-untranslatedregion,187bp3′-untranslatedregionand1137bpopenreadingframe(ORF),whichencodes378aminoacidswithcharacteristicsignalpeptidesandactivationpeptides.ThepepsinogenofS.chuatsialsohashighlyconservedaminoacidresi

5、duesessentialforcatalyticactivityandconformationalmaintenance,whicharetwoessentialaspartylresiduesintheactivesiteandsixcysteineresiduesinvolvedinformingthreedisulphidebonds.ThehomologyofaminoacidsequencesbetweenpepsinogenofS.chuatsiandothervertebratepepsinogensrangesfrom59.9%to91.2%,wh

6、ichindicatesthepepsinogengeneishighlyconservedinevolution.TheNJphylogenetictreeofvertebratesfromtheaminoacidssequencesofpepsinogenshowedallfishwereclusteredasagroup,thepepsinogenofS.chuatsiwasthemostclosetopepsinogenAformⅡaofwinterflounder.ThesuccessfulcloningofpepsinogengenefromS.chua

7、tsinotonlylaysthefoundationforfurtherstudyofitstemporalandspatialexpression,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