牛ME1基因cDNA编码序列的克隆与分析

在NCBI中GenBank里查询已登录的牛的ME1 mRNA序列(GenBank Accession:XM-613987),发现其1-69 bp序列与已知的人、猪和小鼠的ME1基因mRNA序列没有任何相似性,因此,认为这段序列有误。研究利用人的ME1基因mRNA序列作为电子探针共找到44段牛的相关ESTs序列,然后利用此ESTs重叠群拼接成的序列设计了三对引物。提取牛的肝脏和肌肉总RNA,从中克隆测序得到M1为525 bp、M2为1039 bp和M3为1171bp的三段序列,拼接成长度为2015 bp的序列。此段序列与前述ESTs重叠群一致序列完全相同,并与人、猪和小鼠的ME1基因mRNA序列相似性分别达89%、85%和84%,从而证实了本序列的正确性。本序列已在NCBI登录(GenBank Accession:FJ495084)。研究为进一步研究牛的ME1基因结构提供了真实的序列信息。

Cloning and Bioinformatic Analysis of cDNA Encoding Sequence of Bovine ME1 Gene

ME1 gene mRNA sequence was checked from GenBank of NCBI(GenBank Accession: XM-613987),any similarity of its 1-69 bp sequence with ME1 gene mRNA sequence of human,mouse and pig in GenBank could not be found,therefore,this sequence must have something wrong.The present study is using the accessed ME1 gene mRNA sequences of human as electron probe,to find out 44 ESTs related sequences of cattle,and then designed three pairs of primers based on this ESTs contig.Extracted total RNA from liver and muscle of cattle,then through cloning and sequencing,three segments were obtained that there were M1(525 bp),M2(1039 bp) and M3(1171 bp),and splicing them as a 2015 bp sequence.This sequence is identical with the ESTs contig consensus.Additionally,the similarity of this sequence in cattle ME1 gene mRNA is 89%,85% and 84% compared with corresponding sequences in human,pig and mouse,respectively.Therefore,the correctness of this sequence was confirmed.This sequence had accessed to NCBI(GenBank Accession: FJ495084).Our present study will provide more correct sequence information of bovine ME1 gene mRNA for further study on ME1 gene structure in cattle.