蚯蚓纤溶酶基因的cDNA克隆及其序列分析

蚯蚓纤溶酶基因的cDNA克隆及其序列分析 Cloning and Sequence Analysis of Earthworm Fribrinolytic Enzyme Gene cDNA 胡燕 HU Yan 1 2 first-author 孟小林 MENG Xiao-lin 1 2 徐进平 XU Jin-ping 1 2 张俊杰 ZHANG Jun-jie 1 2 王健 WANG Jian 1 2 鲁伟 LU Wei 1 2 武汉大学生命科学学院 武汉大学生命科学学院 College of Life Sciences, Wuhan University College of Life Sciences, Wuhan University 武汉大学生命科学学院,湖北,武汉,430072 武汉大学生命科学学院,湖北,武汉,430072 从粉正蚓成体中提取总RNA,以寡核苷酸Oligo（dT）20为引物合成cDNA第一条链;用RT PCR的方法扩增蚯蚓纤溶酶的cDNA,将其克隆到pUCm T载体上进行DNA序列测定.结果表明,所克隆的蚯蚓纤溶酶cDNA长度为747bp,其中编码区段为738bp,共编码246个氨基酸.将该序列与GenBank中其它的4种蚯蚓纤溶酶序列（LumbricusrubellusEFE III 1、Lumbricusrubelluslumbrokinase 3（1）precursor、Lumbricusbimastuslumbroki nase、Lumbricusrubelluslumbrokinase 1T4precursor）相比,同源性分别为99%,97%,89%,88%;与同源性最高的序列相比,本序列有7个碱基、3个氨基酸与该序列不同. Earthworm fribrinolytic enzyme （EFE） cDNA was amplified by RT-PCR from adult worm of Lumbricus rubellus, and then cloned into pUCm-T vector. Sequence analysis revealed that the cDNA was 747 base pair, and that the encoding fragment was 738 base pair which encoded a protein of 246 amino acid residues. Comparison with four other EFE cDNA sequences （Lumbricus rubellus EFE-III-1、Lumbricus rubellus lumbrokinase-3（1） precursor、Lumbricus bimastus lumbrokinase、Lumbricus rubellus lumbrokinase-1T4 precursor） registered in GeneBank shows 99%,97%,89%,88% homology in DNA sequence respectively. There are seven base substitutions between the reported sequence and the Lumbricus rubellus EFE-III-1, those of which subsequently leads to change of three amino acids. 蚯蚓纤溶酶 cDNA RTPCR 序列分析 earthworm fibrinolytic enzyme cDNA RT-PCR sequence analysis Q785 2004-02-01 2021-04-01 2