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1、鱖胃蛋白酶原基因cDNA全長的克隆與序列分析吳雪峰，趙金良（上海水產大學農業(yè)部水產種質資源與養(yǎng)殖生態(tài)重點開放實驗室，上海200090）關鍵詞：鱖；胃蛋白酶原；cDNA末端快速擴增；克??；序列分析中圖分類號：S917文獻標識碼：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資助項目：上海市重點學科建設項目(Y1
