凡纳滨对虾P23蛋白基因的筛选、表达和生物信息学分析

Screening,cloning and bioinformatics analysis of P23 gene in Litopenaeus vannamei

克隆凡纳滨对虾极端体重个体的组织差异表达基因P23基因并进行生物信息学分析,为进一步研究该基因的功能以及凡纳滨对虾的分子选育等研究提供信息。以凡纳滨对虾雌虾极端体重个体腹部肌肉为实验材料,利用抑制消减杂交(SSH)技术构建极大体重雌性个体和极小体重雌性个体腹部肌肉组织正反向消减cDNA文库:正库(以极大体重个体为试验组,以极小体重个体为驱动组,L-S)和反库(以极小体重个体为试验组,以极大体重个体为驱动组,S-L)消减cDNA文库,并采用实时定量技术分析P23基因的组织表达规律,利用生物信息学对其功能和结构进行预测。以β-actin为看家基因检测两个文库的消减效率分别为210和25,实时定量技术分析结果表明P23基因在体重的调节中起上调作用。P23基因编码区序列全长495bp,编码165个氨基酸,GenBank登录号:JF806619。生物信息学分析发现P23蛋白部分序列含有强疏水性,无跨膜螺旋结构,两种信号肽预测都显示P23含有信号肽。

Screening, cloning and bioinformatics analysis of P23 gene differently expressed in large female and small female of Litopenaeus v,annamei. Suppression subtractive hybridization （SSH） approach was used to isolate differ- ently expressed genes in abdominal muscle samples of the female-shrimp between large female （LF; body weight〉 90 percentile of weight distribution curve） and small female （SF; body weight〈10 percentile of weight distribution curve）. Real time reverse transcriptase-polymerase chain reaction （RT-PCR） was used to analysis the change of P23 mRNA level between large female and small female, and biotechnology were employed to predict gene specific structure and potential motif domain. The subtraction efficiency was estimated by a housekeeping gene β-actin, and the results showed that β--actin was subtracted efficiently at 2^10 and 2^5 folds for SF and LF subtracted cDNA library respectively. RT-QPCR showed that the P23 has an effect to up-regulated expression in weight gain. Whole length of the P23 gene from Litopenaeus vannamei was 495bp （186 amino acid residues）. The sequence has been accessed in GenBank （JF806619）. Partial sequence of the P23 gene had strong hydrophobicity but with no transmembrane helices structure. The signal peptide prediction in P23 gene showed that there was a signal peptide with strong amino acid residues in the partial Sequence of the P23 gene.