布莱凯特牛心脏脂肪酸结合蛋白基因的序列测定及分析

Sequencing and Analyzing of Heart Fatty Acid-binding Protein (H-FABP) Gene in BLACK Cattle

根据GenBank发表的牦牛心脏脂肪酸结合蛋白(heart fatty acid-binding protein,H-FABP)基因的序列设计13对引物,分13段扩增出布莱凯特牛H-FABP基因,采用PCR产物直接测序的方式对各个片段进行测序,使用生物信息学软件对各个片段的测序结果进行拼接,得到布莱凯特牛H-FABP基因的全序列,并对其进行序列分析。结果表明,布莱凯特牛的H-FABP基因(GenBank登录号:FJ756345)是由4个外显子(73、173、102和54 bp)和3个内含子(3463、1892和1494 bp)组成;布莱凯特牛与牦牛、普通牛、山羊、猪、马、人、大鼠、小鼠和鸡等9个物种之间的核苷酸同源性大小依次为99.3%、99.0%、96.3%、92.5%、89.8%、88.3%、83.3%、82.8%、75.6%,其氨基酸的同源性为99.2%、99.2%、96.2%、93.2%、91.7%、88.7%、86.5%、86.5%、77.4%;系统发生树将这些物种总体上分成2支,鸡为独立的一支,布莱凯特牛、牦牛、普通牛、山羊、猪、马、人、大鼠、小鼠为另一独立的大分支。因此,渤海黑牛H-FABP基因具有很强的保守性,其进化树符合物种进化规律。

According to the heart fatty acid-binding protein（H-FABP） gene sequence of yak from GenBank,13 pairs of primers were designed to amplify the whole gene sequence.The H-FABP gene of BLACK cattle was amplified,sequenced and analyzed.The nucleotide sequence was detected and the amino acid sequence of this gene was deduced.Then we compared to deduce amino acid sequence with the homologous H-FABP gene of other species.A phylogenetic tree of the H-FABP gene of BLACK cattle and other species was constructed with MEGA 4.0.The results showed that H-FABP gene of BLACK cattle（FJ756345） had 4 exons（73,173,102 and 54 bp） and 3 introns（3463,1892 and 1494 bp）.After comparison,the identities of the nucleotide sequence of BLACK cattle H-FABP gene with that of yak,general cattle,goat,pig,horse,human,rat,mouse and chicken were 99.3%,99.0%,96.3%,92.5%,89.8%,88.3%,83.3%,82.8%,75.6%,respectively,and the identities of the amino acid sequence of BLACK cattle H-FABP gene with that of yak,general cattle,goat,pig,horse,hu-man,rat,mouse and chicken were 99.2%,99.2%,96.2%,93.2%,91.7%,88.7%,86.5%,86.5%,77.4%,respectively.The phylogenetic results showed that these species were classified into 2 branches as a whole,in which the chicken were on one branch and BLACK cattle,yak,general cattle,goat,pig,horse,human,rat and mouse were another branch.The H-FABP gene of BLACK cattle was highly conserved,the phylogenetic tree of the H-FABP gene accords with the order of evolution.