本文選題：花生　切入點(diǎn)：AhGPAT　出處：《花生學(xué)報》2017年03期

【摘要】：�；D(zhuǎn)移酶基因家族是甘油三酯合成的關(guān)鍵基因。本研究以前期克隆的AhGPAT9與AhLPAAT4基因部分序列設(shè)計引物,構(gòu)建原核表達(dá)載體,轉(zhuǎn)化進(jìn)大腸桿菌BL21,經(jīng)IPTG誘導(dǎo)表達(dá),利用SDS-PAGE檢測表明,重組蛋白在37℃,5h誘導(dǎo)條件下獲得高效表達(dá)。重組蛋白經(jīng)純化和富集,對新西蘭兔進(jìn)行4次免疫,純化獲得的多克隆抗血清,通過間接ELISA檢測,表明獲得了效價比較高的多克隆抗體。通過對重組蛋白純化后樣品進(jìn)行Western Blot分析,結(jié)果顯示在純化樣品相應(yīng)位置有明顯信號,表明所制備的抗體具有很高靈敏度和特異性。并采用制備的抗體對AhGPAT9與AhLPAAT4蛋白在花生不同組織及種子不同發(fā)育時期的表達(dá)進(jìn)行了Western Blot分析。為深入研究AhGPAT9與AhLPAAT4基因的功能提供了科學(xué)數(shù)據(jù)。
[Abstract]:Acyltransferase gene family is the key gene of triglyceride synthesis. In this study, primer was designed based on partial sequences of AhGPAT9 and AhLPAAT4 genes cloned in the early stage, and the prokaryotic expression vector was constructed and transformed into E. coli BL21. The expression was induced by IPTG, and detected by SDS-PAGE. The recombinant protein was highly expressed under induction at 37 鈩，

本文編號：1683618