编码蛋白激酶的人类新基因NEK8的电子克隆

目的：克隆人类基因组中小鼠NEK8基因的同源物。方法：用同源筛选策略，以小鼠NEK8基因作为信息探针，在GenBank数据库中进行TBLASTN分析，将获得的高度同源的EST用VECTORNTI软件拼接成重叠群。利用BLAT分析确定NEK8基因的基因组结构以及染色体定位。利用SMART网上分析工具进行结构域预测，利用Unigene数据库进行表达谱分析。结果：电子克隆到人类NEK8新基因cDNA序列，长度为2858bp，含完整的ORF，编码一条692个氨基酸的多肽，被预测的分子量为74．8KDa，等电点8．04。定位在染色体17q11．2，由15个外显子和14个内含子组成。其编码蛋白含有一个苏氨酸／丝氨酸蛋白激酶结构域，与NEK家族成员有较高的同源性。电子表达谱分析显示NEK8基因在胎盘，胃，结肠，眼，胰腺，骨骼，肾，子宫等组织中表达。结论：电子克隆到编码苏氨酸／丝氨酸蛋白激酶的人类新基因NEK8。

In Silicon Cloning of Human Novel Gene NEK8 Encoding a Protein Kinase

Objective: In silicon cloning of human novel gene ho mology to mouse NEK8. Methods: Using homologous strategy, TBLA STN searched GenBank database with the coding sequence of mouse NEK8 gene. T he ESTs identified were assembled to the contig. BLAT analysis was made by brows ing the UCSC genomic database to determine the gene structure and the mapping in formation. Smart analysis for domain searching. Unigene database was used to stu dy the expression pattner. Results: we cloned a novel human gene NEK8,The length of cDNA is 2858bp, and maped to 17q11.2. It consists of fif teen extons and fourteen introns, encoding a protein with serine/threonine prot ein kinases domain. It is homologous to NEK protein kinase family. And its predi cted molecular weight and isoelectronic point is 74.8KDa and 8.04 respectively . The result in situ blast indicated that NEK8 was expressed in placenta, st omach, cervix, colon, eye, pancreas, bone and kidney. Conclusion: Human novel gene NEK8 encoding a serine/threonine protein kinase was cloned in silicon.