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环链虫草一个含HSF结构域蛋白编码基因的克隆与表达分析

Cloning and Expression of HSF in Cordyceps cateniannulata

  • 摘要:
    目的 明确环链虫草热激转录因子CcHSF759的基本特性及其在环境胁迫响应中的作用,为解析该菌的胁迫耐受机制提供依据。
    方法 基于转录组数据克隆CcHSF759基因,对其进行生物信息学分析(包括序列特征、系统进化、结构域与三级结构模拟),并采用qRT-PCR检测该基因在不同胁迫(高温、低温、高渗、细胞壁干扰剂及氧化胁迫)下的表达模式。
    结果 克隆获得CcHSF759基因,其编码序列(Coding sequence, CDS)全长1005 bp,编码334个氨基酸,预测蛋白分子量为36.52 kDa,理论等电点为6.05;系统进化分析表明其与玫烟色棒束孢、爪哇虫草菌和球孢白僵菌亲缘关系最近;该蛋白具有典型热激因子(Heat shock protein, HSF)结构域、双螺旋结构域及C端低复杂性区域;三级结构以无规则卷曲和α螺旋为主。表达分析显示:在不同胁迫下,CcHSF759CcHSP78CcHSP90CcHSP100 3个热激蛋白(Heat shock protein, HSP)基因呈现胁迫类型依赖性的差异化表达关联,其中CcHSF759CcHSP90在高温、高渗及细胞壁干扰胁迫下呈稳定负向关联,而与其他HSP成员的协同或抑制关系则随胁迫类型和浓度发生动态变化。
    结论 CcHSF759可能参与环链虫草对多种胁迫的转录响应,并可能与部分HSP基因形成胁迫类型依赖的表达关联,其调控功能及分子机制尚需深入验证。

     

    Abstract:
    Objective Characteristics and roles played in response to abiotic stresses of the heat shock transcription factor HSF in Cordyceps cateniannulata were studied.
    Method Based on the transcriptome database, CcHSF759 was cloned using the local BLAST method. Bioinformatics analyses including sequence characterization, phylogenetic analysis, domain prediction, and tertiary structure modeling were conducted on the cloned gene with the expressions under various imposed stresses detected by RT-qPCR.
    Result The successfully cloned CcHSF759 had a CDS of 1005 bp and encoded 334 amino acids with a molecular weight of 36.52 kDa and an isoelectric point of 6.05. Phylogenetically, it closely related to C. javanica, Beauveria asiatica, and B. bassiana. The protein contained typical N-terminal HSF, middle coiled-coil domains, and two low-complexity regions at the C-terminus. Its tertiary structure was composed mainly of random coils and α-helices. Under high or low temperature, high osmotic pressure, cell wall interference, and oxidative stress, CcHSF759, as well as CcHSP78, CcHSP90, and CcHSP100, exhibited differential stress-specific expressions. CcHSF759 and CcHSP90 were consistently inversely correlated to the treatments of high temperature, hyperosmotic condition, and interference to cell wall integrity. Whereas other HSP members displayed variations in their reaction to the abiotic stresses.
    Conclusion CcHSF759 might participate in the transcriptional response and displayed expressions to abiotic stresses different from other HSP proteins in C. cateniannulata. Further study to decipher the regulatory functions and molecular mechanism associated with the heat shock transcription factors is in order.

     

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