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《鱖胃蛋白酶原基因cdna全長(zhǎng)的克隆與序列分析》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫(kù)。
1、鱖胃蛋白酶原基因cDNA全長(zhǎng)的克隆與序列分析吳雪峰，趙金良（上海水產(chǎn)大學(xué)農(nóng)業(yè)部水產(chǎn)種質(zhì)資源與養(yǎng)殖生態(tài)重點(diǎn)開(kāi)放實(shí)驗(yàn)室，上海200090）關(guān)鍵詞：鱖；胃蛋白酶原；cDNA末端快速擴(kuò)增；克??；序列分析中圖分類號(hào)：S917文獻(xiàn)標(biāo)識(shí)碼：ACloningandsequencingofthefull-lengthcDNAofpepsinogengenefromtheMandarinfish，SinipercachuatsiWUXue-Feng,ZHAOJin-Liang(KeyLaboratoryofAquaticGeneticResourcesandAquacu
2、lturalEcosystemCertificatedbytheMinistryofAgriculture,ShanghaiFisheriesUniversity,Shanghai200090,China)Abstract:Pepsinogenisonekindofgastricdigestiveproteinasesbelonglingtoafamilyofasparticproteinases,whichissynthesizedinthegastricmucosaofvertebratesandconvertedtopepsinsintheac
3、idicenvironmentofgastricjuice.Themandarinfish(Sinipercachuatsi)isatypicalcarnivorousfishwhichonlypreyonsmalllivefishesafterhatching.Inordertounderstandthemolecularcharactersofpepsinogeninfish,thefull-lengthcDNAofpepsinogengeneofS.chuatsiwasclonedbymeansofRT-PCRandrapidamplifica
4、tionofcDNAends(RACE)andsequenced.Theresultrevealsa1367bpcDNAfull-lengthsequencecontaining43bp5′-untranslatedregion,187bp3′-untranslatedregionand1137bpopenreadingframe(ORF),whichencodes378aminoacidswithcharacteristicsignalpeptidesandactivationpeptides.ThepepsinogenofS.chuatsials
5、ohashighlyconservedaminoacidresiduesessentialforcatalyticactivityandconformationalmaintenance,whicharetwoessentialaspartylresiduesintheactivesiteandsixcysteineresiduesinvolvedinformingthreedisulphidebonds.ThehomologyofaminoacidsequencesbetweenpepsinogenofS.chuatsiandotherverteb
6、ratepepsinogensrangesfrom59.9%to91.2%,whichindicatesthepepsinogengeneishighlyconservedinevolution.TheNJphylogenetictreeofvertebratesfromtheaminoacidssequencesofpepsinogenshowedallfishwereclusteredasagroup,thepepsinogenofS.chuatsiwasthemostclosetopepsinogenAformⅡaofwinterflounde
7、r.ThesuccessfulcloningofpepsinogengenefromS.chuatsinotonlylaysthefoundationforfurtherstudyofitstemporalandspatialexpression,butalsoprovidesnewmaterialforthemolecularcharacteristicandevolutionoffishpepsinogen.Keywords:Sinipercachuatsi;pepsinogen;rapidamplificationofcDNAends;clon
8、ing;sequenceanalysis收稿日期：2007-07-19資助項(xiàng)目：上海市重點(diǎn)學(xué)科建設(shè)項(xiàng)目(Y1
