陆川猪H-FABP基因克隆与序列分析

Gene cloning and sequence analysis of heart fatty acid- binding protein(H-FABP) gene from Luchuan pigs

为了研究心型脂肪酸结合蛋白(H-FABP)基因与陆川猪肌内脂肪沉积的相关性,寻找猪脂肪沉积的主要候选基因,试验对陆川猪H-FABP基因进行了克隆与序列分析,参照Gen Bank已经公布的猪H-FABP基因序列(EF619344)设计引物,扩增克隆了陆川猪H-FABP基因的编码区序列,并应用DNAStar、Prot Scale等生物软件对其进行了序列分析和蛋白质二级结构预测。结果表明:陆川猪H-FABP基因编码区全长402 bp,编码133个氨基酸,碱基序列在235位点由A突变为G,并引起相应氨基酸由精氨酸转变为甘氨酸。其与野猪的亲缘关系最近,碱基同源性为99.3%;与人类、绵羊和家牛的碱基同源性也在90%以上。陆川猪H-FABP成熟肽包含有25.56%的α螺旋、12.03%的β转角、25.56%的无规卷曲和36.84%的延长线。说明H-FABP基因可作为研究陆川猪脂肪沉积的候选基因之一。

To study the correlation between heart fatty acid - binding protein （ H - FABP） gene and intranluseular fat deposition in Luchaun pig to look for the main candidate genes for fat deposition, the H - FABP gene from Luchaun pigs was used for cloning and sequence analysis. The primers were designed according to pig H - FABP gene sequence published in GeneBank （Accession No. EF619344 ）, and then the sequence of the coding region of the H - FABP gene was also amplified and cloned. The DNAstar and ProtScale and other biological softwares were used to analyze of H - FABP gene sequence and predict the secondary structure of protein. The results showed that the full length of the H - FABP gene from Luchaun pigs was 402 bp, encoding for a protein of 133 amino acids. It was found that the base sequence of the H - FABP gene had a mutation from A to G at 235 site, which caused the corresponding amino acid changing from arginine to glycine. The Luchuan pig had the closest relationship with wild boar, the H - FABP gene of Luchuan pig shared 99.3% homology of base sequence with that of the wild boar, and also shared above 90% of base sequence with that of Homo sapierts, Ovis aries and Bos taurus. The mature peptide of Luchuan pig H - FABP contained α - helix of 25.56%, 13 - corner of 12.03%, random coil of 25.56%, and extension cords of 36.84%. The results indicate that the H - FABP gene can be used as one of the candidate genes for studing the fat deposition of Luchuan pigs.