人乳腺组织中乳铁蛋白基因cDNA的克隆与序列分析

通过RT-PCR技术从人乳腺癌组织中扩增出长度为2259 bp的人乳铁蛋白cDNA,PCR产物经凝胶回收纯化后,克隆在pMD 18-T载体的T位点。测序结果和序列分析显示,与GenBank中收录的人、小鼠、奶牛和山羊的乳铁蛋白cDNA序列的同源性分别为99．73％,82．19％,81．79％和87．28％。其中与人乳铁蛋白cDNA 序列相比,发现有5个碱基因等位基因间的突变而发生变异,1个碱基发生了未知的突变,表明已成功的扩增、克隆了人乳铁蛋白基因cDNA序列,此基因序列的GenBank登录号为AY165046。

Cloning and Sequence Analysis of Human Lactoferrin cDNA

Human lactoferrin (hLF) cDNA fragment about 2 259 bp nucleotide acids in length was obtained through RT-PCR from human mammary cancer tissue. The PCR product was cloned into pMD 18-T vector. The fragment was sequenced and analyzed. The gene homology of the fragment obtained in this study was 99. 73%,82. 19%,81. 79%,and 87. 28% respectively comparable to that of the reported lactoferrin cDNA sequence of human, mouse, cow, and goat in GenBank. The Sequence was aligned with human lactoferrin cDNA in GenBank,and the result suggested there were five allele mutation sites and one unrecognized mutation site. So the human lactoferrin cDNA was cloned successfully,and the sequence number in GenBank was AY165046.