中国农业科学2012，Vol.45Issue(4)：774-785,12.DOI:10.3864/j.issn.0578-1752.2012.04.019
鸭腺苷琥珀酸裂解酶基因序列特征及表达与肌肉肌苷酸含量的相关性分析
Analysis of Sequence Characters of ADSL Gene and Correlation between Gene Expression and IMP Content in Duck
摘要
Abstract
[Objective] The CDS (coding sequences) of duck ADSL gene was cloned. The study was aimed to analyze gene sequences, protein structure, relative expression levels of ADSL gene in muscle and the correlation between gene expression and inosine monophosphate (IMP) content in ducks. This research revealed the ADSL gene structure of excellent native ducks of China, established a foundation for further research of its biological function and relationship with flavor substances. [Method] Reverse transcriptase PCR was employed to clone the cDNA of ADSL gene. The coding region sequences, protein structure and phylogenetic relationships were analyzed by bioinformatics software. Real-time-PCR was employed to examine the relative expression levels of ADSL gene in muscle of ducks. IMP content in breast and leg muscle was detected by HPLC. [Result] The CDS of ADSL gene inducks had 1380 bp in length, encoded 459 amino acid residues and showed 94.77% sequence identity with chicken. The physical and chemical properties of duck ADSL indicated that ADSL is a basic protein. And the protein has more strongly signal peptide and transmembrane segments in cells. The secondary structure of duck ADSL was mainly made by a helixes. Phylogenetic analysis showed that duck and chicken were in the same clustering, and then clustered with human, mice and other mammals, both of which had a distant relationship with fish and amphibian. The expression levels of ADSL gene among different varieties were extremely different (P<0.01), that between female and male were extremely different (P<0.01), and that among different locations were different (P<0.05) except those in cherry valley ducks (P>0.05). IMP content among different varieties were extremely different (P<0.01), and among different sexes and locations were very different (P<0.05). [Conclusion] The CDS of ADSL gene in ducks showed a high sequence identity with chicken. The expression levels of ADSL gene and the IMP content of relative locations were extremely different (P<0.01) or different (P<0.05) among different varieties, sexes and locations, respectively. ADSL gene relative expression and IMP content were significantly correlated.关键词
鸭/ADSL/克隆/mRNA表达/肌苷酸/高效液相色谱法Key words
duck/adenylosuccinate lyase/cloning/mRNA expression/IMP/high-performance liquid chromatography引用本文复制引用
徐善金,虞德兵,汪峰,何宗亮,张剑锋,于海龙,杜文兴..鸭腺苷琥珀酸裂解酶基因序列特征及表达与肌肉肌苷酸含量的相关性分析[J].中国农业科学,2012,45(4):774-785,12.基金项目
教育部高校博士点基金(B200928)、江苏省自然科学基金(SBK201021501)、江苏省农业综合开发产业化经营项目(320111291)、南京市科技发展计划项目(2010sd210013) （B200928）
