摘要
目的：大鼠ZA73基因(GenBank accession number：AF011363)是本实验室从大鼠支气管上皮恶性转化细胞模型中采用mRNA差异显示技术克隆到的EST片段，与支气管上皮细胞恶性转化相关。根据此EST序列，克隆人全长基因。材料与方法：运用生物素标记探针筛选人cDNA文库、将筛选基因进行cDNA快速终末端扩增(Rapid amlification of cDNA ends)，直至获得全长序列。结果：HRNT-1 cDNA总长4256 bp，开放阅读框架2760 bp，5′非编码序列253bp，3′非编码序列1240 bp，已被GenBank收录于Nr数据库中(Accession number：AF223393)。结论：采用生物素标记cDNA文库筛选和cDNA快速终末端扩增相结合的技术是克隆全长cDNA快速而有效的方法，尤其适用于那些长度超过2kb的基因。
Objective： An EST fragment, Rat ZA73 (GenBank Accession Number： AF011363 ), was cloned by using mRNA differential display technique from rat tracheal epithelial (RTE) neoplastic transformed cell model induced by α particles radiation. Homology search in GenBank showed that rat ZA73 was a novel EST. Based on the previous work, this study is to identify human full length sequence of EST. Methods： According to the sequence of rat ZA73, a biotin labeled probe was used to screen a human cDNA library, and then the gene sequence was extended by RACE (rapid amplification of cDNA ends). Results： Human gene named HRNT 1 (GenBank Accession Number： AF223393) was sequenced. Its total length is 4256 bp in length, with an ORF located in the region between 254 and 3013 bp. 5′ UTS (untranslated sequences) is 253 bp, 3′ UTS is 1240 bp. Conclusions： The combination of biotin labeled screening of cDNA library and RACE is an effective method to clone full length cDNA, especially for those with sequence longer than 2 kb.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2000年第9期939-942,共4页
Chinese Journal of Cancer
基金
本课题受国家重点基础研究专项经费!(编号:G1998051208)资助
国家自然基金!(编号:39870670)资助
