撒坝猪AIDA基因编码区克隆、序列特征分析及组织表达量检测

Cloning,Bioinformatics and Tissue Expression Analysis of AIDA Gene in Saba Pig

对不同猪的AIDA基因编码区进行扩增、测序、拼接,采用生物信息学方法对撒坝猪AIDA编码蛋白进行功能和性质分析,并通过实时荧光定量PCR检测其在撒坝猪不同组织中的表达水平。PCR扩增测序表明:该基因在撒坝猪的编码区序列没有发生错义突变,序列长921 bp,编码306个氨基酸。AIDA蛋白属于亲水蛋白,没有跨膜结构和信号肽,含有两个卷曲螺旋结构,有1个糖基化位点和29个磷酸化位点,无CpG island;二级结构是以无规卷曲为主的混合型蛋白;AIDA基因在大白猪的背最长肌和背脂中表达量要高于撒坝猪,而撒坝猪AIDA基因在胰脏中的表达量最高,其他组织中的表达量依次为脾脏、背脂、肾脏、肺、大脑、肝脏、心脏和背最长肌。

The study was aimed to amplify,sequence and splice the coding region of AIDA gene of different pigs,and the CDS of Saba pigs was analyzed by bioinformatics.The expression levels in different tissues were detected by real-time fluorescence quantitative PCR.PCR sequencing showed that there was no missense mutation in the coding region of the gene,the sequence length was 921 bp,encoded 306 amino acids and it was an instability,hydrophilic protein.AIDA protein had no cross-membrane structure,signal peptide and contained two typical crimp helix structures.The AIDA protein had one glycosylation site and 29 phosphorylation sites,and it had no CpG island.The secondary structure was a mixed protein with random coil.The Real-time PCR results showed that AIDA was the most expressed gene in pancreas,followed by spleen,backfat,kidney,lung,brain,liver,heart and longissimus dorsi.The AIDA gene of Yorkshire was expressed higher than Saba pig in the longissimus dorsi and backfat.