‘合作猪’TDRP1基因的克隆及生物信息学分析

发布时间：2018-11-04 09:29

【摘要】：【目的】丰富猪TDRP1基因研究的基础数据.【方法】以猪NM_001198925序列为参考,设计特异性引物,通过克隆测序获得‘合作猪’TDRP1基因完整CDS序列,并进行生物信息学分析,同时采用实时荧光定量PCR方法检测TDRP1基因在不同组织中的表达特性.【结果】获得了TDRP1基因完整的CDS序列(GenBank登录号:KU743254),共684bp,其编码186个氨基酸多肽.与参考序列相比,在编码区第33、348位点处发生了同义突变(C→G、C→T).TDRP1基因分子式为C_(897)H_(1421)N_(259)O_(287)S_2,理论等电点(PI)为5.86,不稳定系数为62.66,疏水指数为64.03,平均亲水性为-0.939,属不稳定可溶性酸性蛋白质.二级结构以无规则卷曲和α-螺旋为主,属混合类蛋白质.亚细胞定位结果显示,TDRP1编码的蛋白质在遗传物质复制和转录、翻译过程中发挥功能的可能性分别为26.3%、14.3%,明显高于发挥其它功能的可能性.mRNA表达分析表明,TDRP1基因在垂体和睾丸中高表达,肺、肾、小脑、卵巢中度表达,肝、脾、大脑、胃、小肠中低表达,心脏组织中不表达.【结论】成功克隆了‘合作猪’TDRP1基因的完整CDS区序列,并发现了2个SNP位点;多组织转录表达分析表明TDRP1在垂体和睾丸中表达较高,可为深入研究TDRP1基因的功能提供参考.
[Abstract]:[objective] to enrich the basic data of porcine TDRP1 gene research. [methods] using porcine NM_001198925 sequence as reference, specific primers were designed to obtain the complete CDS sequence of 'cooperative pig' TDRP1 gene by cloning and sequencing, and bioinformatics analysis was carried out. Meanwhile, real-time fluorescence quantitative PCR was used to detect the expression of TDRP1 gene in different tissues. [results] the complete CDS sequence of TDRP1 gene (GenBank accession number: KU743254) was obtained, which encoded 186 amino acid peptides. Compared with the reference sequence, a synonymous mutation (C _ (897) H _ (1421) N _ (259) O _ (287) S _ 2) occurred at the 33348 locus of the coding region (C _ (897) H _ (1421) N _ (259) O _ (287) S _ 2). The theoretical isoelectric point (PI) is 5.86, the instability coefficient is 62.66, the hydrophobic index is 64.03, and the average hydrophilicity is -0.939. The secondary structure is mainly irregular curl and 伪-helix, which is a mixed protein. The subcellular localization results showed that TDRP1 encoded proteins were more likely to function in the process of genetic material replication and transcription and translation than other functions, and the mRNA expression analysis showed that the function of the proteins was significantly higher than that of other functions. TDRP1 gene was overexpressed in pituitary and testis, moderately expressed in lung, kidney, cerebellum, ovary, liver, spleen, brain, stomach and small intestine. [conclusion] the complete CDS region of 'Hehe pig' TDRP1 gene was cloned and two SNP loci were found. Multiple tissue transcriptional analysis showed that TDRP1 was highly expressed in pituitary and testis, which could provide a reference for further study on the function of TDRP1 gene.
【作者单位】：甘肃农业大学动物科学技术学院;兰州大学第一医院;甘肃农业大学生命科学技术学院;
【基金】：甘肃省科技重大专项“高繁殖力猪新品系选育研究”(092NKDA036)
【分类号】：S828

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