金黄色葡萄球菌β溶血素基因的克隆与表达

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1、中文摘要金黄色葡萄球菌B．溶血素是金黄色葡萄球菌分泌的外毒索之一。是重要的毒力因子，它可引起多种疾病，奶牛乳房炎的主要致病因子之一就是金黄色葡萄球菌分泌的B．溶血素。奶牛乳房炎是一种多因素疾病，给世界各国奶牛业造成巨大的经济损失。患牛牛奶中体细胞数大幅增加，会降低乳蛋AbstractBeta-hemoIysinwaSoneofthemostimportantextracellulartoxinsproducedbyviruientStrainsof研qp妙幻∞cc淞d“阳琊(s口“理时)，whichwaSanimpon

2、antvirulencef犯torandassociatedwjththeeti0109icalofavarjetyofjnfectiousdiseasesjnanimaIs，includingmastjtisMaStitiswaSakjndofmu】tiplef如tOrdisease，whichresultedinenomouseconomiclossinwo—darIdplayedanimpon锄tro】einthemilkquality．ThesomaticnumbcrofthemiIkofthemastitis

3、ca砌ewasincreaSed，whichcoulddecreasethecontentionofmjlkprotein，1actoseandmilkfat吼Ifantibioticswereusedtotreattheinfectedcanle，whichmight1eadtomuch『emajnDoublep01lutionofthesomaticandantibioticjnfreshmilk，mightbringd枷agetothemilkproductintheaspectofqual时锄dnavoLMas

4、titiswaSap—maryobstacletomedeVelopmentofa11imajhusballdryofChina，foran廿biotictreatlnentSaffbctedthemilkqualjtyseriously'thusvaccinationmightbeawaytopreventt}1edisease．Asaresult，todevelop廿1eengineeringVaccinemightbeawaytosolVe也eproblem．As仃ainof最4“旭淞SlOusedi11this

5、researchwasgained行om廿1emastitismilks锄ples．Thegeneencodingbeta．hemolysinof脚砂，DcDcc埘口舯埘was啪plifiedbyPolymerasechainReaction(PcR)锄dwaslinkedtot11eph缸)-18一TtoconsⅡucttbeB-hemolysincIoneplaSmid．ThecompleteDNAsequenceofthecloned8-hemolysingene疗Dm＆矽砂kDcc蚶日“旭淞wassequenc

6、酣aI】dconfi姗edbyDNAsequencea11alysis．Theresultsshowed廿1att}leDNAsequencehom0109ywaSmoret11an99．2％aftercomparingw油sequencepublishedinGenbank，锄dt11ehomologyoftheencodingproteinwas100％，A11tllisassuredthatthesequenceoftlle且-hemolysinof飘础yfococc淞口“雕淞hadhi曲conserv撕on锄d

7、homolog弘Thent}IerecombinationwaSdoubleen巧meinciteda11dinsertedimopET-32-aexpressionvector．TheexpressionofrecombinarItbeta-hemolysinproteinwasdetcctedbySDS母AGEaIldWestemBlot．Theresultsshowed出atthemolecuIarweightoffusionproteinofbeta小em01ysinwasabout57l(Da，锄dexpre

8、ssionlevelw鹊30％．TheexpressionofrecombinalltproteinwaSpurined，then也eproductwasusedin血etreadmjlltest，thereSultsshowedt11att11eexpressionpmteinalsocou】dmakethesheeperyth