Institute of Plant Pathology and Applied Microbiology, Heilongjiang Bayi Agricultural University, Daqing, 163319 Heilongjiang People's Republic of China ; State Key Laboratory of Crop Stress Biology in Arid Regions, Northwest A & F University, Yangling, 712100 Shaanxi People's Republic of China.
Institute of Plant Pathology and Applied Microbiology, Heilongjiang Bayi Agricultural University, Daqing, 163319 Heilongjiang People's Republic of China.
Indian J Microbiol. 2014 Sep;54(3):310-4. doi: 10.1007/s12088-013-0439-3. Epub 2013 Dec 4.
By using EST database from a full-length cDNA library of Curvularia lunata, we have isolated a 2.9 kb cDNA, termed PKAr. An ORF of 1,383 bp encoding a polypeptide of 460 amino acids with molecular weight 50.1 kDa, (GeneBank Acc. No. KF675744) was cloned. The deduced amino acid sequence of the PKAr shows 90 and 88 % identity with cAMP-dependent protein kinase A regulatory subunit from Alternaria alternate and Pyrenophora tritici-repentis Pt-1C-BFP, respectively. Database analysis revealed that the deduced amino acid sequence of PKAr shares considerable similarity with that of PKA regulatory subunits in other organisms, particularly in the conserved regions. No introns were identified within the 1,383 bp of ORF compared with PKAr genomic DNA sequence. Southern blot indicated that PKAr existed as a single copy per genome. The mRNA expression level of PKAr in different development stages were demonstrated using real-time quantitative PCR. The results showed that the level of PKAr expression was highest in vegetative growth mycelium, which indicated it might play an important role in the vegetative growth of C. lunata. These results provided a fundamental supporting research on the function of PKAr in plant pathogen, C. lunata.
利用旋孢腔菌全长 cDNA 文库中的 EST 数据库，我们分离出了一个 2.9kb 的 cDNA，称为 PKAr。该 cDNA 含有一个编码 460 个氨基酸的开放阅读框，分子量为 50.1kDa（GeneBank Acc. No. KF675744）。PKAr 的推导氨基酸序列与Alternaria alternate 和 Pyrenophora tritici-repentis Pt-1C-BFP 的 cAMP 依赖性蛋白激酶 A 调节亚基分别具有 90%和 88%的同一性。数据库分析表明，PKAr 的推导氨基酸序列与其他生物中的 PKA 调节亚基具有相当大的相似性，特别是在保守区域。与 PKAr 基因组 DNA 序列相比，ORF 内的 1383bp 中没有发现内含子。Southern blot 表明 PKAr 作为基因组中的单个拷贝存在。使用实时定量 PCR 检测了不同发育阶段 PKAr 的 mRNA 表达水平。结果表明，PKAr 的表达水平在营养生长菌丝中最高，这表明它可能在旋孢腔菌的营养生长中发挥重要作用。这些结果为 PKAr 在植物病原菌旋孢腔菌中的功能研究提供了基础支持。
