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钟树荣, 金扬, 李东升, 王婉瑜, 熊郁良. 2005: 圆斑蝰蛇毒七个C-型凝集素样蛋白亚基的cDNA克隆与序列分析(英文). 动物学研究, 26(4): 337-343.
引用本文: 钟树荣, 金扬, 李东升, 王婉瑜, 熊郁良. 2005: 圆斑蝰蛇毒七个C-型凝集素样蛋白亚基的cDNA克隆与序列分析(英文). 动物学研究, 26(4): 337-343.
ZHONG Shu-rong, JIN Yang, LI Dong-sheng, WANG Wan-yu, XIONG Yu-liang. 2005. Molecular Cloning and Sequence Analyses of cDNAs Encoding Seven C-type Lectin-like Protein Subunits from Daboia russellii siamensis. Zoological Research, 26(4): 337-343.
Citation: ZHONG Shu-rong, JIN Yang, LI Dong-sheng, WANG Wan-yu, XIONG Yu-liang. 2005. Molecular Cloning and Sequence Analyses of cDNAs Encoding Seven C-type Lectin-like Protein Subunits from Daboia russellii siamensis. Zoological Research, 26(4): 337-343.

圆斑蝰蛇毒七个C-型凝集素样蛋白亚基的cDNA克隆与序列分析(英文)

Molecular Cloning and Sequence Analyses of cDNAs Encoding Seven C-type Lectin-like Protein Subunits from Daboia russellii siamensis

  • 摘要: 按照Promega公司的mRNA提取试剂盒操作手册,从圆斑蝰蛇(Daboia russellii siamensis)的毒腺中提取mRNA;利用RT-PCR的方法进行体外扩增,获得C-型凝集素蛋白的基因,克隆到pMD18-T载体中。随机挑选14个阳性克隆进行核酸测序,获得7个编码不同蛇毒C-型凝集素样蛋白亚基的cDNA,分别命名为DRS-L1、DRS-L2、DRS-L3、DRS-L4、DRS-L5、DRS-L6和DRS-L7。由基因序列推导出的氨基酸序列表明,克隆到的7个蛇毒C-型凝集素样蛋白的亚基中均有糖识别结构域存在。BLAST分析显示,仅有DRS-L1的蛋白序列与目前已知的蛇毒C-型凝集素样蛋白的α亚基相似。序列同源性比较并结合半胱氨酸位点分析,推测DRS-L1和DRS-L2可能分别是圆斑蝰蛇毒Ⅹ因子激活剂的轻链LC2和LC1。 DRS-L3和DRS-L4可能是高分子量的蛇毒C-型凝集素样蛋白的β亚基,而DRS-L5和DRS-L6可能是低分子量的蛇毒C-型凝集素样蛋白的β亚基。DRS-L7可能是类似于血小板膜糖蛋白Ib结合蛋白的β亚基。

     

    Abstract: Total mRNA was extracted from a venom gland of snake Daboia russellii siamensis following the manufacturers protocol of the PolyATtract System 1000 kit purchased from Promega Biotech. cDNAs encoding C-type lectins were amplified by RT-PCR and subcloned into a pMD18-T vector. Fourteen positive clones were selected for nucleotide sequencing and seven cDNAs encoding various snake venom C-type lectin-like protein precursors, designated as DRS-L1, DRS-L2, DRS-L3, DRS-L4 DRS-L5, DRS-L6 and DRS-L7, were obtained. Amino acid sequences of these proteins were deduced and each contains a carbohydrate recognition domain. Of all the deduced protein sequences, only DRS-L1 seemed to represent a closer sequence similarity to α subunits of other known snake venom C-type lectin-like proteins using the BLAST program. Homology comparison combined with analysis of cysteine position indicate that DRS-L1 and DRS-L2 are probably the light chain LC2 and LC1 of factor Ⅹ activator from Daboia russellii siamensis venom, respectively. DRS-L3 and DRS-L4 might be the β subunits of higher molecular weight C-type lectin-like proteins while DRS-L5 and DRS-L6 might be β subunits of lower molecular weight C-type lectin-like proteins. DRS-L7 might be the β subunit of a platelet membrane glycoprotein Ib-binding protein similar to echicetin.

     

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