香猪Noggin基因克隆与序列分析

以贵州榕江香猪为实验材料,设计2对特异性的寡核苷酸引物,RT-PCR扩增了Noggin基因的上下两段序列,将其克隆到T载体上测序,拼接两段序列后得到香猪Noggin基因的完整序列。此序列由896个碱基组成,编码232个氨基酸,前19个氨基酸为信号肽,后213个氨基酸为成熟肽。Genbank blast对香猪Noggin基因编码氨基酸与其他物种进行检索分析:与人、鼠、鸡、爪蟾和东方鲀等氨基酸同源性为54.1%~98.7%。SWISS-MODEL服务器预测了香猪Noggin成熟肽的三维结构,与人和鼠对比:主要由于第99和112位的氨基酸改变而引起三维结构不同。

Cloning and Sequence Analysis of Noggin Gene of Xiang-pig

Noggin shows more and more important to the regulation of growth and development in animal.Designing two pair primers,two fragments of up and down were obtained through RT-PCR.The whole gene of Noggin of Xiang pig was made up by the two arrayed fragments and the length of full Noggin sequence is 896 bp.The deduced ORF amino acid sequence contained a putative signal peptide of 19 amino acids and a mature peptide of 232 amino acids.Genbank blast analysis shows that Noggin amino acid sequence resulted in Xiang-pig has 54.1%~98.7% similarity scores with human,mouse,chick,xenopus and fugu rubripes.The protein structure of Noggin of xiang pig was predicted by SWISS-MODEL.The result demonstrated that the difference of structure compared with human and mouse is due to the amino acid of matural peptide substitution at 99th and 112th.