橘青霉全局性调控因子Pci-veA 基因的克隆与分析
Molecular Cloning and Bioinformatic Analysis of the Global Regulator Pci-veA in Penicillium citrinum
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摘要: 克隆和分析橘青霉 Penicillium citrinum的全局性调控因子Pci‐veA 基因．设计简并引物，用PCR扩增获得Pci‐veA 基因的核心片段，利用RACE和TAIL‐PCR技术对核心片段的3’和5’末端进行延伸，将获得的3个片段进行拼接并设计引物，克隆得到完整的 Pci‐veA 基因．结果表明：Pci‐veA 基因全长1774 bp ，包含一个70 bp的内含子，cDNA序列为1704 bp ，编码567个氨基酸；Pci‐veA 与多种已报道的真菌veA 基因具有较高的同源性，且含有 veA基因特定的双组份核蛋白定位信号区NLSI和Pat7定位信号区域．这些结果为进一步研究 P. citrinum中美伐他汀生物合成的全局调控机理奠定了基础．Abstract: To clone the global regulator Pci‐veA of Penicillium citrinum .Based on the conserved sequence region of gene veA reported from fungus ,the core fragment of gene Pci‐veA was cloned from P .citrinum mycelial samples by using degenerate primers amplify method .And ,the 3'cDNA and 5'cDNA ends of Pci‐veA were amplified using RACE and TAIL‐PCR methods .The full‐length sequence of Pci‐veA gene is 1 774 bp ,w hich contains an intron with length of 70 bp and an open reading frame (ORF) encoding a pro‐tein with 567 amino acids .Pci‐veA shows highly homologue with several reported veA genes ,and contains specific motifs of bipartite nuclear localization signal motif NLSI and pat 7 motif .These results suggest that Pci‐veA will be useful for understanding of molecular regulation mechanism of mevastatin biosynthesis in P .citrinum .
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