摘要
为克隆和筛选肝癌特异表达基因.应用抑制消减杂交技术对肝癌及癌旁组织进行消减杂交,产物PCR扩增后,进行克隆.共得到1820个克隆,1776个克隆有100～800 bp的插入片断,阳性率达98%,说明人肝癌组织特异表达cDNA消减文库构建成功,同时我们对cDNA克隆进行测序,证明这些克隆的功能是和肝癌的发生高度相关的.抑制消减杂交是克隆特异表达基因的有效方法.
In order tO generate a differentially express genes cDNA library of human hepatocellular carcinoma. The cDNA fragments of differentially expressed genes between hepatocellular carinoma （HCC） tissue and paracancerous liver tissues were isolated by suppression subtractive hybridization （SSH）. After amplificated with PCR, the cDNA fragments were cloned. As the result, the SSH library, we obtained, contained 1820 positive clones. 98 percent of pos- itive clones were 100-800 bp insert-fragments. The sequence analysis of 260 cDNA fragments has proved that these fragments were tighly correlated with the occurrence and development of the hepatocellular carcinoma. These results suggest a high quality and high representative cDNA library of human hepatocellular carcinoma was constructed successfully.
出处
《南开大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第4期115-118,共4页
Acta Scientiarum Naturalium Universitatis Nankaiensis
基金
天津市科委重点基金资助项目(003803411)
关键词
抑制消减杂交
肝癌
基因文库
CDNA克隆
suppression subtractive hybridization （SSH）
hepatocellular carcinoma （HCC）
cDNA library
cDNA clone
