祝氏麻鸡（Gallus gallus）终端父本CAB39L基因克隆和生物信息学及组织表达分析

柴睿堂; 祝春三; 李和鸣; 黄永欢; 杨秀荣

doi:10.13417/j.gab.042.000393

祝氏麻鸡（Gallus gallus）终端父本CAB39L基因克隆和生物信息学及组织表达分析

Cloning,Bioinformatic and Tissue Expression Analysis of CAB39L Gene in Zhushi Partridge Chicken（Gallus gallus）

摘要

摘要: 为了探究钙结合蛋白样39(calcium binding protein 39 like,CAB39L)的蛋白结构及其基因的组织表达规律，本研究以祝氏麻鸡(Gallus gallus)终端父本为试验材料，克隆CAB39L基因并进行生物信息学分析和组织表达分析。通过克隆获得CAB39L基因的CDS(conding sequence)全长序列，应用实时定量PCR检测CAB39L基因在祝氏麻鸡终端父本各组织中的表达水平。结果表明：祝氏麻鸡终端父本CAB39L基因的编码区长度为1 005 bp，共编码334个氨基酸；与参考序列(索引号：NM＿001006272)进行序列比对，存在两处同义突变(c.852G>T, c.874T>C)。物种间同源性分析显示，祝氏麻鸡终端父本CAB39L基因与原鸡(Gallus gallus)、雉鸡(Phasianus colchicus)、中国家鹅(Anser cygnoiders domesticus)、鸽(Columba livia)、人(Homo sapiens)、黑猩猩(Pan troglodytes)、家鼠(Mus musculus)、猴子(Macaca mulatta)、斑马鱼(Danio rerio)CAB39L基因的相似性分别为99.8%、97.5%、93.7%、92.6%、83.6%、83.6%、83.5%、82.5%、75.4%；系统进化树分析显示，祝氏麻鸡终端父本与原鸡的亲缘关系最近，其次是与雉鸡的亲缘关系比较近，与斑马鱼的亲缘关系最远，这表明CAB39L基因在禽类和非禽类之间有明显的遗传距离。CAB39L蛋白的等电点为7.69，为碱性蛋白；亲疏水性分析显示该多肽链氨基酸序列的平均亲疏水性为-0.320，为亲水蛋白；蛋白功能结构域预测结果显示该蛋白只含有Mo25结构域，且不属于跨膜蛋白和分泌蛋白。实时定量PCR结果显示，CAB39L基因在睾丸中的表达量最高，这表明CAB39L基因可能与公鸡睾丸组织的生长发育或功能有关，进而影响公鸡的生长情况。本研究结果为进一步研究鸡CAB39L基因的生物功能奠定了理论基础。

Abstract: In order to explore the protein structure of calcium binding protein 39 like（CAB39L） and the tissue expression of its gene,the male parent of the terminal of Zhushi Partridge chicken（Gallus gallus） was used as experimental material to clone the CAB39L gene and conduct bioinformatics analysis and tissue expression profile analysis.The CDS（conding sequence） sequence of CAB39L gene was cloned,and the expression level of CAB39L gene was detected by real time quantitative PCR.The results showed that the coding region of CAB39L gene was 1 005 bp in length,encoding 334 amino acids.Compared with the reference sequence（accession number:NM＿001006272）,there were two synonymous mutations（c.852G＞T,c.874T＞C）.Analysis of homology among species revealed that the similarity between the CAB39L gene of Zhushi Partridge chicken and the CAB39L gene of jungle fowl（Gallus gallus）,pheasant（Phasianus colchicus）,chinese domestic goose（Anser cygnoiders domesticus）,rock pigeon（Columba livia）,human（Homo sapiens）,chimpanzee（Pan troglodytes）,house mouse（Mus musculus）,monkey（Macaca mulatta） and zebrafish（Danio rerio） were 99.8%,97.5%,93.7%,92.6%,83.6%,83.6%,83.5%,82.5% and 75.4%,respectively.Phylogenetic tree analysis showed that Zhushi Partridge chicken was most closely related to the jungle fowl,more closely related to pheasant,and most distantly related to zebrafish.It indicated that CAB39L gene had a significant genetic distance between poultry and non-poultry.The theoretical isoelectric point of CAB39L was 7.69,which was a basic protein.Hydrophilic analysis showed that the average hydrophilic hydrophobicity of the amino acid sequence of CAB39L was-0.320,which was a hydrophilic protein.Functional domain prediction showed that the protein contained only Mo25 domain and did not belong to transmembrane proteins or secretory proteins.Real time quantitative PCR results showed that CAB39L gene expression was the highest in testis,indicating that CAB39L gene may be related to the growth,development or function of rooster testicular tissue,and thus affect the growth of rooster.Our results provide a theoretical basis for further studies on the biological function of chicken CAB39L gene.

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