乌苏里蝮蛇具抑制血小板聚集活性的金属蛋白酶分离纯化及基因信息学分析

发布时间：2018-06-04 11:03

  本文选题：金属蛋白酶 + 解整连蛋白　； 参考：《吉林大学》2005年硕士论文

【摘要】：提取中国长白山乌苏里蝮蛇毒腺mRNA,逆转录合成cDNA,并以其为模板,进行PCR 扩增,PCR 产物克隆入pGEM-T Easy 载体并进行了序列分析。结果显示,乌苏里蝮蛇前金属蛋白酶原基因开框读码序列由1434bp 组成,编码478 个氨基酸。由核苷酸顺序推导的氨基酸序列可以看出,前金属蛋白酶原基因最初的翻译产物是酶原前体;依次含有18 氨基酸组成的信号肽,171 个氨基酸组成的酶原区和由289 个氨基酸组成的前金属蛋白酶原基因(200 个氨基酸组成的金属蛋白酶结构域、16 个氨基酸组成的间隔区和73 个氨基酸组成的解整合蛋白结构域)。乌苏里蝮蛇前金属蛋白酶原的金属蛋白酶结构域含有3 对二硫键;解整合蛋白结构域含有6 对二硫键和特征性RGD(精氨酸-甘氨酸-天冬氨酸)结构。其基因序列和结构域组成与GenBank 中蛇毒金属蛋白酶/解整合蛋白呈现高度同源性属于PⅡ。氨基酸序列blast 比对发现,酶原区和解整链蛋白结构域呈现极高的同源性,而金属蛋白酶结构域却出现了极高的变异,推测这些变异结构区是为了适应不同的底物、不同受体或同一受体的不同结构域。
[Abstract]:MRNAs were extracted from the venom gland of Agkistrodon halys in Changbai Mountain, China, and were synthesized by reverse transcription. The PCR products were amplified by PCR and cloned into pGEM-T Easy vector and sequenced. The results showed that the open frame reading sequence of prometalloproteinase gene of Agkistrodon halys was composed of 1434bp and encoded 478 amino acids. The amino acid sequence deduced from nucleotide sequence shows that the original translation product of prometalloproteinase gene is proenzyme precursor. An 18-amino acid signal peptide composed of 171 amino acids in turn and a prometalloproteinase gene consisting of 289 amino acids, composed of 200 amino acids, composed of 16 amino acids. Domain and 73 amino acid composition of the unintegrated protein domain. The metalloproteinase domain of Agkistrodon halys Pallas has 3 pairs of disulfide bonds and 6 pairs of disulfide bonds and characteristic RGD (arginine glycine aspartic acid) structure. Its gene sequence and domain composition were highly homologous to the snake venom metalloproteinase / integrin in GenBank, belonging to P 鈪，

本文编号：1977126