阿部鲻(鱼叚)(鱼虎)P-gp蛋白编码区基因克隆与分析  

Cloning and sequential analysis of P-gp in Mugilogobius abei

作　　者：程章[1] 聂湘平[1] 王芳[2] 李凯彬[2] 

机构地区：[1]暨南大学水生物研究中心,广州510632 [2]中国水产科学院珠江水产研究所,广州510380

出　　处：《安全与环境学报》2009年第1期4-8,共5页Journal of Safety and Environment

基　　金：科技部公益研究专项资助项目(2004DIB1Z-029);NSFC-广东联合基金项目(U0633006)

摘　　要：利用RT-PCR(Reverse Transcription Polymerase Chain Reaction)分离阿部鲻(Mugilogobius abei)P-gp(P-glycoproteins)CDS(Sequence coding for amino acids in protein)基因,并进行序列测定与分析。结果表明,cDNA全长2 073 bp,其中阅读框1 958 bp,编码652个氨基酸,估算的蛋白质相对分子质量为71.87 kD;5′非翻译区115 bp。序列同源性分析表明,不同进化地位动物的P-gp编码序列存在高度同源性,显示P-gp基因在系统进化上高度保守。生物信息学分析表明,阿部鲻P-gp基因不含有编码蛋白信号肽的序列,预测结果显示,有4个跨膜区,编码蛋白含ATP结合盒(the ATP-binding cassette)内膜转运typ-1型结合域。This paper is aimed to present our research and cloning of P-gp CDS cDNA from the liver of Mugilogobius abel by using RT- PCR. For our purpose, we have designed pairs of primers Ppl and Pp2 in accordance with the sequence of fish M. abel domain. At the same time, we have also laid out the other three gene-speeific primers Ppsl, Pps2 and Pp3 in accordance with the sequence of 3＇ end cDNA. As is known, P-glycoproteins （P-gps） has been developed for a number of years to confer the multi-drug resistant phenotypes to tumor cells or tumors of the invalids by preventing cellular accumulation of drugs whereas the transmembrane P-gps help to mediate the efflux of a large number of structurally and functionally diverse, moderately hydrophobic compounds, including endogenous regulators, such as glucocorticoids, drugs and other natural products. However, the sequential analysis has revealed that the CDS cDNA sequence contains 2073 bp, 1 958 bp open, and the frame （ORF）, the structure of which was encoded by 652 amino acids. Therefore, the deduced amino acid sequence aligned with those of P-gp genes from different species also display high degree of sequential homology as is shown by the phylogenic tree. Moreover, the bio-informatics analysis indicates that the M. abel P-gps do not contain the code protein signal peptide. Such analysis has thus provided a new kind of material that may help to indicate the P-gp gene molecular evolution and the similar comparison results. But the forecast results from the analysis demonstrate that the protein can be divided into four crisscross membrane areas while the code protein contains the ATP-binding cassette transporter integral membrane of type-1 fused domain, which proves the great significance for further studies of the structure and function of its code protein as well as the formation of specific antibody. Therefore, the reaction of P-gp in the fish to the exposure of various xenobioties can thus be used as a potential biomarker for monitoring environmental pollution.

关 键 词：水生生物学 阿部鲻鰕鯱 P-GP CDS 基因克隆 核苷酸序列分析 

分 类 号：X171.5[环境科学与工程—环境科学]