CAJ | 학술논문

猪肺炎支原体（Mhp）是猪气喘病的主要病原体。利用MhpNJ株基因通过SOE-PC（splicingwithoverlapextensionPCR）R扩增和突变，获得了目的片段并插入表达载体pET-28a（＋）中，然后转入宿主菌BL21（DE3）。重组质粒经1mmol／LIPTG在37℃温度下诱导5h，目的蛋白可达到总蛋白的14．1％。WesternBlotting证明表达的重组蛋白具有猪肺炎支原体反应原性，用该重组蛋白建立的ELISA方法可检测到猪肺炎支原体抗体。此为该病基因工程疫苗抗原的筛选和ELISA诊断试剂盒的研制奠定基础。
저폐염지원체（Mhp）시저기천병적주요병원체。이용MhpNJ주기인통과SOE-PC（splicingwithoverlapextensionPCR）R확증화돌변，획득료목적편단병삽입표체재체pET-28a（＋）중，연후전입숙주균BL21（DE3）。중조질립경1mmol／LIPTG재37℃온도하유도5h，목적단백가체도총단백적14．1％。WesternBlotting증명표체적중조단백구유저폐염지원체반응원성，용해중조단백건립적ELISA방법가검측도저폐염지원체항체。차위해병기인공정역묘항원적사선화ELISA진단시제합적연제전정기출。
Mycoplasma hyopneumoniae （Mhp） is the etiologic agent of mycoplasma pneumonia in swine. In this study the DnaK gene of strain NJ is amplified by gene splicing with overlap extension PCR using a pair of inside-mutation primers. The PCR product is inserted into expression plasmid pET-28a （＋）. The recombinant plasmid is transformed into competent cells of BL21（DE3） and it contains correct object fragments by sequencing. It is induced to express by 1 mmol/L IPTG at 37 ~C for 5 hours. The dose of the recombinant protein is nearly 14. 1~ of the total product. The expression product of interest and its biological activities are characterized with Western blotting and ELISA analysis. This study provides the basis for diagnose agent and future preparation of new vaccine a~ainst Mvconlasma hvonneurnnniae.