摘要
LRP16是我们应用甲基化敏感的限制性界标基因组扫描 (restrictionlandmarkgenomicscanning ,RLGS)技术时发现的一个与白血病复发相关的新基因。为钓取这一新基因的全长cDNA ,本实验采用了cDNA末端快速扩增法 (rapidamplificationofcDNAend ,RACE)。通过对RACE法若干步骤进行优化 ,克隆得到了LRP16基因cDNA的 5′ 与 3′ 非翻译区 ,进而获得其全长及完整的开放阅读框架 ,并作为新基因被GenBank收录。上述结果表明 ,RACE技术是钓取未知基因全长cDNA敏感而快速的方法 ,尤其是对 5′ 及 3′
LRP16 is a novel gene which was found in our laboratory by using methylation-sensitive restriction landmark genomic scanning(RLGS) technique. In order to clone the full-length cDNA of this leukemia relapse associated gene, the method of rapid amplification of cDNA end (RACE) was employed. By optimizing some procedures of RACE method, the 5′-and 3′-untranslated region of LRP16 cDNA was successfully sequenced. Then, the full length of LRP16 cDNA and open reading frame(ORF) was constructed and was registered in GenBank. The above-mentioned procedure demonstrated RACE technique is a rapid and sensitive method for cloning unknown gene. Especially, it is very useful to cloning the 5′- and 3′-untranslated region of a novel gene.
出处
《中国实验血液学杂志》
CAS
CSCD
2001年第1期18-21,共4页
Journal of Experimental Hematology
基金
国家自然科学基金资助 编号 3 9970 82~~
