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Expression of Acidic Phospholipase A2-1 from Guangxi Ophiophagus Hannah (King Cobra) Venom
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- Author:
- No author available
- Journal Title:
- JOURNAL OF MODERN CLINICAL MEDICAL BIOENGINEERING
- Issue:
- 1
- DOI:
- 10.3760/cma.j.issn.1674-1927.2003.01.002
- Key Word:
- 眼镜王蛇毒;酸性磷脂酶A2;表达;复性;活性测定
Abstract： 将广西眼镜王蛇毒酸性磷脂酶A2-1(APLA2-1)基因克隆至表达载体pET28a(+),转化入大肠杆菌BL21,经过诱导,SDS-PAGE检测,重组质粒pET28a-APLA2-1在18KD有一明显的表达条带,表达产物APLA2-1约占细菌总量30%,并以包涵体的形式存在,将产物变复性后用Sephadex G-75 纯化,产物经过SDS-PAGE检测只有单一条带,通过Western-blot检测,证实纯化产物是酸性磷脂酶A2.对表达的APLA2进行酶活性和药理活性的测定.至此我们在大肠杆菌中表达了广西眼镜王蛇毒APLA2,这将为以后对PLA2的结构与功能的研究打下了良好的基础.
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