【摘要】 以与东北马鹿物种最近的牛(Bos taurus cattle,序列号为AB076403)的肌肉生长抑制素(myo-statin,MSTN)基因序列为模版,根据该基因保守序列进行引物设计,扩增产物连接入pMD18-T载体,克隆出3个外显子片段。根据5′-GU-AG-3′规则和同源基因比对拼接出带有部分5′、3′末端的cDNA,并采用生物信息学技术对编码氨基酸序列进行蛋白质同源性比较和二级、三级结构等分析。结果表明:所获序列全长为1 128 bp,编码375个氨基酸;具有较明显的螺旋、片层和无规卷曲等二级结构;有1个较明显的跨膜区和1个疏水区,并发现明显的信号肽和糖基化位点。三级结构同源建模预测结果显示与Human Activin A的相似性为42%。结果说明东北马鹿和其他哺乳类生物的肌肉生长抑制素基因之间关系密切,并同属于TGF-β家族。更多还原

【Abstract】 The three exon fragments of the myostatin(MSTN)gene of Manchurian wapiti were cloned from the latest species of cattle(Bos taurus cattle accession number: AB076403) for the templates,with primers according to the conservative of MSTN gene sequencing.The products of PCR were linked into pMD18-T vector and then cloned three exons.MSTN gene containing partial of the 5′and 3′end of the cDNA was spliced according to 5′-GU-AG-3′ rules and homologous matching.The bioinformatic analysis was used to predict amino acid homology alignment,the secondary and tertiary domain and so on.The extended sequence is composed of 1 128 base pairs coding 375 amino acids.The secondary structure prediction showed that the alpha helix,bata sheet and coil.One transmembrane areas and one hydropathicity profiles were found,signifi-cant signal peptide and glycosylation sites were also found.The three diamensions structure has been gained by the way of mock construction of molecular confirmation which showed the 42% homology with the Human Activin A.It showed that the close relationship between Manchurian wapiti and other mammals for genes of the myostatin and the myostatin gene is belong to the family TGF-β.The present paper could accumulate data useful data for inter-species homogeneous gene cloning by PCR and the study on structures and functions of myostatin(MSTN) gene.更多还原