日本百脉根牻牛儿基牻牛儿基氢化酶的电子克隆及进化分析

根据物种间同源基因相对保守的特点，利用生物信息学的方法，以拟南芥牻牛儿基牻牛儿基氯化酶（GGH）基因cDNA序列为信息探针，搜索GenRank中的HTGs数据库，找到一个与之高度匹配的日本百脉根基因组DNA序列LJT46M09，用GENSCAN软件分析该序列得到一个包含2个外显子和1个内含子的基因。以此基因序列BLAST检索GenBank中的est-others数据库获得了同源的EST片段。进行EST拼接后得到该基因的cDNA序列，GenBank登录号为DQ013361，由1389bp核苷酸组成，具有完整的开放阅读框架（ORF），推测编码蛋白为462个氨基酸。推导的氨基酸序列与大豆、截形苜蓿、烟草、拟南芥、小麦、水稻的牻牛儿基牻牛儿基氢化酶基因氨基酸序列的一致率分别为91％、89％、84％、81％、73％、74％。访基因与叶绿素等的生物合成有关。

In Silico Cloning and Evolution Analysis of Geranylgeramyl Hydrogenase Gene from Lotus corniculatus var. Japonicus

With the development of EST projects and bio-informatics, in silico cloning has been a new strategy of gene cloning. With Arabidopsis thaliana GGH cDNA sequence as a querying probe to search the Lotus corniculatus vat. japonicus HTGs database, a highly homologous DNA sequence, which was referred as LjT46M09 clone, was obtained, The result of gene prediction with performing GENSCAN program showed that this gene contained 2 exons and 1 intron. With this predicted sequence to BLAST GenBank est-others database, many homologous EST sequences, which can be edited a unique sequence, was obtained, which was the same as the predicted sequence from genomie DNA. The full-length genc of 1 389 bp contained a complete ORF, encoding 462 amino acids. Its GenBank accession number was DQ013361. The deduced amino acid showed 91%, 89%, 84%, 81%, 73% and 74% homology with GGH from Glycine max, Medieago truncatula, Nicotiana tabacum, Arabidopsis thaliana and Triticum aestivum, respectively. This eDNA sequence was confirmed with EST sequences.