摘要
从兔脾脏细胞中分离提取总RNA ,经反转录PCP(RT -PCR)扩增出兔巨嗜细胞阳离子多肽 (MCP- 1)cDNA ,插入经EcoRI和XbaI双酶切的pUC19中 ,构建了克隆质粒pUCDEF .进行了限制性酶切鉴定和序列分析 ,结果在扩增出的cDNA 2 88个碱基中 ,在前片段中有一个碱基与发表的兔MCP - 1cDNA序列不同 ,即第 157位碱基由G变为A ,导致编码的氨基酸由丙氨酸变为苏氨酸。该cDNA全长 2 88bp ,编码 94个氨基酸 ,由编码信号肽 。
In order to expression the cDNA of rabbit macrophage cationic peptide-1(MCP-1)in eukaryotic cell,total RNA were isolated from Rabbit spleen cells and the cDNA encoding macrophage cationic peptides(MCP-1)was amplified by the reverse transcription PCR(RT-PCR).The cDNA was inserted into vectors pUC19 and the cloning vector pUCDEF was constructed.The cDNA was identified by restriction endonucleases and sequenced.It was found to differ in nucleotide as compared to the cDNA sequence (Ganz, et al .1989).This changes cause amino acid substitution in the propiece sequence at position 157 where a Ala changed into a Thr.This cDNA encodes 94 amino acids whose length is 288bp,including sequence encoding signal peptide,propiece and mature peptide.
出处
《生物技术》
CAS
CSCD
2000年第5期3-6,共4页
Biotechnology
基金
吉林省自然科学基金资助项目(吉科合字第963509号)
