中文摘要：

目的 构建广西眼镜王蛇毒酸性磷脂酶A2-1（APLA2-1）在不同载体的重组表达质粒，在Ecoli中表达APLA2-1并比较不同表达系统对APLA2-1的表达效果。方法将广西眼镜王蛇毒酸性磷脂酶A2-1（AP-LA2-1）基因克隆至表达载体pBLMVL：和pET28a（＋），分别转化人大肠杆菌RRl和BL21，经过诱导表达，应用SDS-聚丙烯酰胺凝胶（SDS-PAGE）及Western blot观察重组蛋白表达情况。结果 成功构建了重组质粒pBLMVLz-APLA2-1和pET28a-APLA2-1。pBLMVL2-APLA2-1在SDS-PAGE上没见明显表达带，在Westernblot上可见一14kD的表达带。pET28a-APLA2-1在SDS-PAGE上有一明显的18kD表达条带，表达产物AP-LA2-1约占细菌总量30％，并以包涵体的形式存在。结论 APLA2-1可在大肠杆菌中表达，pET28a（＋）对APLA2-1的表达效果优于pBLMVL2。

英文摘要：

Objective To construct recombinant plasmids encoding acidic phospholipase A2-1 （APLA2-1） from Guangxi Ophiophagus hannah （King cantor） Venom and express in E. coli so as to compare the expression efficiency in different expression systems. Methods A cDNA encoding acidic phospholipase A2-1 （APLA2-1） from Guangxi Ophiophagus hannah （King can- tor） Venom was cloned into bacterial expression vector pBLMVL2 ,pET28a（q-） and expressed in E. coli RR1 ,BL21 respectively. The expressed recombinant proteins were analyzed by sodi- um docedyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE） and Western blot. Results The recombinant plasmids pBLMVLz-APLA2-1 and pET28a-APLA2-1 were constructed sucessfully. No obviously expression band was observed in SDS-PAGE by using pBLMVL2 expression system, but one band with molecular weight 14 kDa was observed in Western blot. Moreover, a strong band with molecular weight 18 kDa was observed in SDS-PAGE and Western blot using pET28a（＋） expression system. The expression amount of APLA2-1 was about 30M of that of total cell proteins. In addition, the protein was produced as insoluble inclusion bodies. Conclusion The APLA2-1 could be expressed in E. coli. The results showed that there was better expression efficiency in pET28a（＋） than pBLMVL2.