克隆鲁西斗鸡三重基序蛋白62(Chicken triple motif protein 62,TRIM62)基因并对其编码产物进行生物信息学分析,检测其在不同组织和器官中的表达情况。以鲁西斗鸡肺脏cDNA为模板,克隆TRIM62基因序列,运用生物信息学软件对其编码蛋白的亲/疏水性、跨膜螺旋结构、修饰结构、结构域、高级结构和互作蛋白网络等进行预测分析。采用qRT-PCR技术检测TRIM62基因在鲁西斗鸡不同组织中的表达情况。鲁西斗鸡TRIM62基因CDS区为1428bp,编码475个氨基酸,编码蛋白分子式为C2410H3773N675O712S22,分子量为54。30kD,等电点(PI)为6。40,无信号肽,不存在跨膜区;存在2个明显的亲水区,其编码不稳定的亲水性蛋白。TRIM62蛋白由α-螺旋(39。16%)、β-转角(4。21%)、无规则卷曲(37。68%)和延伸链(18。95%)组成。qRT-PCR结果显示,TRIM62mRNA在鲁西斗鸡肺脏中表达量最高,在胰腺、腺胃中的表达量次之,在肌肉组织中表达量最低。本研究成功克隆了鲁西斗鸡TRIM62基因,并初步研究了其组织表达规律,结果为TRIM62基因编码蛋白功能的深入研究奠定了基础,为鲁西斗鸡品种保护及开发利用提供有价值的参考资料。
Bioinformatics and Tissue Expression Analysis of TRIM62 Gene in Luxi Game Chickens
The Luxi game chicken triple motif protein 62(TRIM62)gene was cloned and its coding product was analyzed by bioinformatics,and its expression levels in different tissues and organs were detected.The cDNA of Luxi game chicken lung was used as a template to amplify the TRIM62 gene sequence.Bioinformatics software was used to predict and analyze the hydrophilicity/hydrophobicity,transmembrane helix structure,modified structure,domain,advanced structure and interaction protein network of the encoded protein.The expression of TRIM62 gene in different tissues of Luxi game chickens was detected by qRT-PCR.The CDS region of TRIM62 gene in Luxi game chickens was 1 428 bp,encoding 475 amino acids.The molecular formula of the protein encoded by TRIM62 gene was C2410H3773N675O712S22,the molecular weight was 54.30 kD and the isoelectric point(PI)was 6.40,and there was no signal peptide and transmembrane region.There are two obvious hy-drophilic regions,and the encoded protein is unstable and is a hydrophilic protein.TRIM62 protein was composed of α-helix(39.16%),β-turn(4.21%),random coil(37.68%)and extended strand(18.95%).The results of qRT-PCR showed that TRIM62 mRNA in Luxi game chic-kenswas expressed with the highest expression in lung,followed by pancreas and glandular stomach,and the lowest expression in muscle.In this study,the TRIM62 gene in Luxi game chickens was successfully cloned,and the expression pattern of TRIM62 was preliminarily stu-died.The results of this study laid a foundation for the further study of the function of chicken TRIM62 gene encoded protein.
Luxi game chickenTRIM62 genebioinformaticstissue expression