药用真菌杨树菇钙调素基因及活性初步鉴定

The Molecular Cloning and Bioactivity Characterization of Calmodulin from Medicinal Fungus Agrocybe aegerita

摘要: 目的：克隆杨树菇抗肿瘤相关蛋白基因，初步鉴定其活性。方法：构建cDNA文库，随机测序得到杨树菇钙调素(calmodulin，CaM)基因，并对其进行生物信息学分析，通过检测其在HeLa细胞中表达引起的细胞凋亡及衰老的变化，初步鉴定其抗肿瘤活性。结果：得到全长为588 bp的杨树菇钙调素cDNA序列，编码150个氨基酸，其氨基酸序列在整个钙调素家族中十分保守。钙调素基因在真核细胞中表达没有引起肿瘤细胞的凋亡和衰老。结论：杨树菇钙调素序列具有CaM保守序列模式，是CaM家族的成员。 Objective: The antitumor-associated gene of Agrocybe aegerita was cloned to identify the bioactivity. Method: cDNA library was constructed and the gene calmodulin was obtained by random sequencing. The eukaryotic expression vector was constructed to detect cell apoptosis and senescence activity. Result: The calmodulin gene was cloned with 588 bps code for 150 amino acid residues, and the amino acid sequence was very conserved in calmodulin family. The expression of cloned gene in HeLa cells did not induce the cell apoptosis or senescence. Conclusion: The sequence of calmodulin contained conserved motif and belonged to calmodulin family.