张娟, 虎红红, 邓占钊, 等. 静原鸡AK1基因序列分析及真核表达载体的构建[J]. 华南农业大学学报, 2021, 42(2): 17-25. DOI: 10.7671/j.issn.1001-411X.202006037
    引用本文: 张娟, 虎红红, 邓占钊, 等. 静原鸡AK1基因序列分析及真核表达载体的构建[J]. 华南农业大学学报, 2021, 42(2): 17-25. DOI: 10.7671/j.issn.1001-411X.202006037
    ZHANG Juan, HU Honghong, DENG Zhanzhao, et al. Sequence analysis of AK1 gene from Jingyuan chicken and construction of its eukaryotic expression vector[J]. Journal of South China Agricultural University, 2021, 42(2): 17-25. DOI: 10.7671/j.issn.1001-411X.202006037
    Citation: ZHANG Juan, HU Honghong, DENG Zhanzhao, et al. Sequence analysis of AK1 gene from Jingyuan chicken and construction of its eukaryotic expression vector[J]. Journal of South China Agricultural University, 2021, 42(2): 17-25. DOI: 10.7671/j.issn.1001-411X.202006037

    静原鸡AK1基因序列分析及真核表达载体的构建

    Sequence analysis of AK1 gene from Jingyuan chicken and construction of its eukaryotic expression vector

    • 摘要:
      目的  探究宁夏地方品种静原鸡前期转录组测序筛选出的AK1基因编码蛋白的结构与功能,构建其真核表达载体。
      方法  根据GenBank上已公布的原鸡AK1基因序列,针对其CDS区设计特异性引物,通过克隆测序对AK1基因CDS区SNPs进行快速筛查,构建AK1基因的真核表达载体,并进行编码区生物信息学功能分析。
      结果  AK1基因编码区全长585 bp,编码194个氨基酸;AK1蛋白无跨膜结构域,存在2个CpG岛,18个磷酸化位点,10个抗原表位,为稳定的水溶性蛋白,空间结构以α–螺旋和无规则卷曲为主。亚细胞定位结果显示,AK1蛋白主要位于细胞质内;GO富集分析发现,AK1基因同样富集在细胞质中,与亚细胞定位结果一致。基因共表达分析发现,在与AK1互作的基因中,AK1AMPD1PKM2基因存在共表达,共表达系数分别为0.116和0.063。成功构建出AK1-pEGFP-N1载体。
      结论  该研究结果为后续AK1蛋白功能以及AK1作为肌苷酸相关基因的深入研究提供了科学依据。

       

      Abstract:
      Objective  To explore the structure and function of protein encoded by AK1 gene screened by transcriptome sequencing of Ningxia local breed Jingyuan chicken, and construct its eukaryotic expression vector.
      Method  According to the original chicken AK1 gene sequence published on GenBank, specific primers were designed for its CDS region, and the CDS region SNPs of AK1 gene were screened quickly by cloning and sequencing. The eukaryotic expression vector of AK1 gene was constructed. The bioinformatics function of the coding region was analyzed.
      Result  The full length of AK1 gene coding region was 585 bp, which encoded 194 amino acids. AK1 protein had no transmembrane domain, and it had two CpG islands, 18 phosphorylation sites and 10 antigenic epitopes. AK1 protein was a stable water-soluble protein, and the spatial structure was mainly α-helix and irregular crimp. The results of subcellular localization showed that AK1 protein was mainly located in the cytoplasm, and GO enrichment analysis showed that AK1 gene was also enriched in the cytoplasm, which was consistent with the result of subcellular localization. Gene co-expression analysis of genes interacting with AK1 showed that AMPD1 and PKM2 coexpressed with AK1 gene, and the co-expression coefficients were 0.116 and 0.063, respectively. The AK1-pEGFP-N1 vector was successfully constructed.
      Conclusion  The results of this study provide a scientific basis for further study of the function of AK1 protein and AK1 as an inosinic acid-related gene.

       

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