PCR cloning of type II polyhydroxyalkanoate biosynthesis genes from two Pseudomonas strains.
PCR cloning of type II polyhydroxyalkanoate biosynthesis genes from two Pseudomonas strains.
复制标题
PCR 克隆来自两种假单胞菌菌株的 II 型聚羟基脂肪酸酯生物合成基因。
DOI：
10.1111/j.1574-6968.2001.tb10637.x
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发表时间：
2001
影响因子：
2.1
通讯作者：
G. Chen
中科院分区：
文献类型：
--
作者：
G. Zhang;X. Hang;P. Green;K. Ho;G. Chen
利用假单胞菌ⅱ型pha位点的特性，采用PCR克隆策略克隆了假碱性假单胞菌HBQ06中的phaC1和硝基还原假单胞菌0802中的phaC2两个多羟基烷酸合成酶基因。从PCR产物中鉴定出phaC1 （P. nitroreducens HBQ06）和phaC2 （P. nitroreducens 0802）的完整开放阅读框（orf）。利用序列信息，直接从基因组DNA中克隆出完整的PHA合成酶基因，并通过傅里叶变换-红外光谱和气相色谱法证实其在大肠杆菌中表达。分析了PhaC1和PhaC2蛋白之间的差异，认为这两种蛋白具有不同的功能和进化历史。
Two polyhydroxyalkanoate synthase genes, phaC1 from Pseudomonas pseudoalcaligenes HBQ06 and phaC2 from Pseudomonas nitroreducens 0802, were cloned using a PCR cloning strategy based on the type II pha loci property of Pseudomonas strains. The complete open reading frames (ORFs) of phaC1 (P. nitroreducens HBQ06) and phaC2 (P. nitroreducens 0802) were identified from the PCR products. Using the sequence information, the complete PHA synthase genes were PCR cloned directly from the genomic DNA and expressed in Escherichia coli as confirmed by Fourier transform-infrared spectroscopy and gas chromatography. The differences between PhaC1 and PhaC2 were analyzed and the two proteins were suggested to contain different functions and evolution history.