艾美球虫线粒体加工肽酶β亚基编码基因的分离、表征及表达
Isolation, characterization, and expression of the gene encoding the beta subunit of the mitochondrial processing peptidase from Blastocladiella emersonii.
作者信息
Costa Rocha C R, Lopes Gomes S
机构信息
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo 05599-970, Brazil.
出版信息
J Bacteriol. 1998 Aug;180(15):3967-72. doi: 10.1128/JB.180.15.3967-3972.1998.
Abstract
A 2.3-kb BamHI-KpnI fragment was isolated from a partial genomic library and shown by nucleotide sequence analysis to contain the entire coding region of the gene encoding the beta subunit of the Blastocladiella mitochondrial processing peptidase (beta-MPP). The predicted beta-MPP protein has 465 amino acids and a calculated molecular mass of 50.8 kDa. S1 nuclease protection assays revealed an intron, 209 bp in size, interrupting the coding region between the putative signal sequence and the mature protein. Northern blot analysis showed that beta-MPP mRNA levels decrease significantly during B. emersonii sporulation, reaching basal levels in the zoospore stage. The amount of beta-MPP protein, determined in Western blots, unlike its mRNA, does not vary significantly throughout the fungal life cycle.
摘要
从一个部分基因组文库中分离出一个2.3kb的BamHI - KpnI片段，通过核苷酸序列分析表明，该片段包含编码芽枝霉线粒体加工肽酶β亚基（β - MPP）的基因的完整编码区。预测的β - MPP蛋白有465个氨基酸，计算分子量为50.8kDa。S1核酸酶保护试验显示有一个大小为209bp的内含子，打断了推定信号序列和成熟蛋白之间的编码区。Northern印迹分析表明，在艾美球虫孢子形成过程中β - MPP mRNA水平显著下降，在游动孢子阶段达到基础水平。在蛋白质免疫印迹中测定的β - MPP蛋白量，与其mRNA不同，在整个真菌生命周期中没有显著变化。
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