日本百脉根γ-生育酚甲基转移酶基因的电子克隆及进化分析  

In silico Cloning of Gamma-tocopherol Methyltransferase Gene from Lotus corniculatus var.japonicus

作　　者：孟宪萍[1] 李晓晓[1] 李雅轩[1] 蔡民华[1] 胡英考[1] 

机构地区：[1]首都师范大学生命科学学院,北京100037

出　　处：《安徽农业科学》2006年第24期6457-6459,共3页Journal of Anhui Agricultural Sciences

基　　金：北京市教委科技发展计划项目基金资助(200310028112)

摘　　要：根据物种间同源基因相对保守的特点,利用生物信息学方法,以拟南芥-γ生育酚甲基转移酶cDNA序列为信息探针,在GenBank的HTGs数据库中找到与之高度同源的日本百脉根基因组DNA序列——LjT44D21克隆。通过GENSCAN分析及序列拼接得到日本百脉根"γTMT基因的cDNA序列,GenBank的登陆号为DQ013360。用该序列对GenBank中的est-others进行相似性搜索,获得了44条日本百脉根的EST序列,经拼接后发现与基因组预测的基因序列一致,表明该基因是真实存在和表达的。该cDNA序列长为1 077 bp,具有完整的开放阅读框架(1～1 077 bp)。推测编码蛋白为358个氨基酸,该蛋白序列与大豆、水稻、小麦、拟南芥、油菜、苜蓿和衣藻等物种的-γTMT蛋白质序列进行了同源性比较分析,具有很高的同源性。In silico cloning is a new method for gone cloning. A rabidopsis thidiana γ-TMT eDNA sequence （GenBank accession number AY090280） being used as a querying probe to BLAST the HTGs database, a highly homologous DNA sequence, which was referred as LjT44D21elone, was obtained. After performing the GENSCAN program, a positive gene, which comprised of 6 exons and 5 introns, was found. With the predicted sequence to BLAST GenBank est-others database, 44 ESTs homologous sequences hit can be edited to a unique sequence which was the same as the predicted sequence from genomic DNA.These results showed that this gene authentically existed and expressed. This 1 077 bp gene contained a complete ORF （1 bp-1 077 bp）, encoded 358 amino acids, which was very conserved with Glycine max, Oryza sative, Triticum aestivum, A rabidopsis thaliarta, Brassica oleraceo, Medicago truncatula and Chlamydomonas reinhardtii, respectively.

关 键 词：日本百脉根 Γ-生育酚甲基转移酶 电子克隆 EST 

分 类 号：Q94-333[生物学—植物学]