快速克隆SET家族新成员SET07

Quick cloning of a novel SET family member,SET07

目的:本实验基于生物信息学资源,快速克隆获得SET基因家族新成员SET07,并预测其相关生物学性质,为进一步探讨该基因的功能奠定基础。方法:以含有SET结构域的EST片段为基础,电子延伸获得含有SET结构域的新的编码基因,通过RT-PCR技术从胎脾组织中克隆得到该基因的cDNA序列,利用Northern印迹技术证实SET07基因的全长,并利用生物信息学工具预测其相关生物学性质。结果:获得新的SET家族成员,命名为SET07基因,证实该基因全长为3388bp。分析发现其开放阅读框架为1114bp,编码347个氨基酸,相对分子质量38×103,等电点9.57,在羧基端含有SET结构域,与H4-K20甲基转移酶同源。结论:上述结果提示SET07基因可能具有甲基转移酶的功能,在调节细胞生长周期、控制肿瘤的发生发展中发挥重要作用。

Objective: To quickly clone a novel SET family member, SET07,on the basis of bioinformatic resources,and to predict the biochemical characteristics of the gene in order to lay a foundation for the further research on its function. Methods: 381bp of SET domain was extended by AutoCTG tool based on dbEST database. A new gene containing SET domain was obtained. Its cDNA was cloned from fetal spleen tissue by RT-PCR and sequenced. The full length of SET07 was tested by Northern blot. The biochemical characteristics of SET07 were predicted by software from ExPASY and EMBL. Results: A novel SET family member was obtained by means of bioinformation methods. The coding sequence of the gene was cloned from fetal spleen tissue and named SET07. Northern blot proved that mRNA of SET07 was about (3 388 bp.) Its ORF ranged from +12 to +1 055,encoding 347 amino acids, with SET domain in C terminus. The gene may be ubiquitously found in all kinds of tissues.SET07 protein is about 38×10^(3),with pI 9.57 and a transmembrane helices in 6-26.It lacks signal peptide. SET07 is highly identical to H4-K20. Conclusion:The results suggest that SET07 functions as a histone methyltransferase which plays an important role in the life cycle of cells and carcinogenesis.