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Gene Cloning and Sequence Analysis of a Hybrid Acidic Phospholipase A2 from Guangxi OphiophagusHannah (King Cobra) Venom
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- Author:
- No author available
- Journal Title:
- JOURNAL OF MODERN CLINICAL MEDICAL BIOENGINEERING
- Issue:
- 6
- DOI:
- 10.3760/cma.j.issn.1674-1927.2002.06.002
- Key Word:
- 眼镜王蛇毒;磷脂酶A2;基因;克隆
Abstract： 目的研究广西眼镜王蛇毒杂合性酸性磷脂酶A2(APLA2)的基因克隆与序列分析.方法从广西眼镜王蛇毒腺中提取总RNA,根据种属关系接近的台湾眼镜蛇毒APLA2两端非翻译区的保守序列设计引物,利用RT-PCR进行体外扩增,获得磷脂酶A2基因,克隆至pMD18-T载体,经测序筛选出APLA2-1,APLA2-2及一个新的酸性磷脂酶A2(命名为APLA2-3).根据测序结果确定了APLA2-3基因的全序列,并由此推导编码的氨基酸序列. 结果利用Antheprot 4.3c蛋白质序列分析软件包计算它的等电点为4.885,相对分子质量为16.543kD,并预测了它的二级结构和部分理化性质.同源性比较表明,APLA2-3的1～39位氨基酸残基序列与APLA2-2相同,其同源性为64%,而40～108位氨基酸残基序列与APLA2-1相同,其同源性为69%,109位氨基酸残基序列皆不同于这两种APLA2.结论 揭示了APLA2具有基因多样性,可能与物种进化和生物活性有关,并为进一步研究PLA2的结构与功能的关系提供更多的信息.
- [1]王秋雁,舒雨雁,庄茂辛,等.广西眼镜王蛇毒一种新的酸性磷脂酶A2的基因克隆和性质[J].2001,(6).
- [2]王秋雁,庄茂辛,林文珍,等.广西眼镜王蛇毒酸性磷脂酶A2的分离纯化和性质研究[J].2001,(1).
- [3]刘小龙,钟晓燕,吴祥甫,等.尖吻蝮蛇毒碱性磷脂酶A2的表达及其生化特征[J].2000,(3).
- [4]Liu X,Wu X,Zhou Y.Identification of key residues responsible for enzymatic and platelet-aggregation-inhibiting activities of acidic phospholipase A2S from Agkistrodon halys Pallas.[J].2001,10(1).
- [5]刘小龙,潘华,杨冠珍,等.尖吻蝮蛇毒磷脂酶A2基因的克隆与序列分析[J].1999,(1).
- [6]R M, Kramer,G C, Checani,A, Deykin,etc.Solubilization and properties of Ca2+-dependent human platelet phospholipase A2.[J].1986,878.
- [7]S, Brunie,J, Bolin,D, Gewirth,etc.The refined crystal structure of dimeric phospholipase A2 at 2.5 A. Access to a shielded catalytic center.[J].1985,260(17).
- [8]Liu J,Wu X,Zhong X,etc.Expression, purification and biochemical characterization of a recombinant phospholipase A2, with anticoagulant activity from Agkistrodon halys Pallas.[J].2001,10(1).
- [9]舒雨雁,庄茂辛,周陆生,等.广西眼镜王蛇毒碱性磷脂酶A_2（PLA_2）的分离纯化及其性质的研究[J].1989,(5).
- [10]B W, Dijkstra,R, Renetseder,K H, Kalk,etc.Structure of porcine pancreatic phospholipase A2 at 2.6 A resolution and comparison with bovine phospholipase A2.[J].1983,168(1).