摘要
利用差异显示PCR技术获得的一条在胃癌癌旁和正常组织表达量高的EST W12 3,通过与GenBank的dbest库进行电子杂交 ,选取了与W12 3同源度高的若干EST ,在它们共有的保守序列设计了用于扩增的寡聚核苷酸引物 ,利用cDNA末端快速扩增 (RACE)技术得到了 7条带有polyA尾的 3′EST ,进行序列分析后 ,发现它们均是代表新基因或不同剪接体的EST ,且具有共同的保守序列 ,已登录GenBank。
Previously, W123, a EST(Expression Sequence Tag)fragment derived from normal gastric and paracarcinoma tissue, was cloned using differentiated display PCR technique. A few homologous EST sequences were captured when processing similarity search in human EST database. To search for additional W123 homologous genes in gastric tissue, we designed a primer for 3′ RACE (Rapid Amplification of cDNA End)from highly conserved region among the above ESTs. Using this primer and another provided by 3′ RACE kit, seven EST fragments with poly(A) tail were cloned. Comparison analysis with GenBank demonstrated all these ESTs represented 7 novel genes containing a fragment of common sequence. The results suggested that combinating bioinformatic resource of EST dtabase with 3′ RACE technique is a rapid and effective method for seeking for homologous genes in a specific tissue.
出处
《生物技术通讯》
CAS
2001年第4期257-259,共3页
Letters in Biotechnology
基金
"九五"军队科研基金项目 ( 96M14 5 )
