来自三叶草根瘤菌1536的ce/8A基因的克隆与特性分析
Cloning and characterization of ce/8A gene from Rhizobium leguminosarum bv. trifolii 1536.
作者信息
An J M, Lim W J, Hong S Y, Shin E C, Kim E J, Kim Y K, Park S R, Yun H D
机构信息
Division of Applied Life Science, Gyeongsang National University, Chinju, Korea.
出版信息
Lett Appl Microbiol. 2004;38(4):296-300. doi: 10.1111/j.1472-765x.2004.01485.x.
AIMS
To isolate the cellulase gene from Rhizobium leguminosarum bv. trifolii 1536.
METHODS AND RESULTS
By the shot-gun method a clone (cel8A) harbouring 3.1 kb genomic DNA fragment from R. leguminosarum bv. trifolii 1536 was obtained. The cel8A gene coded 348 amino acids and it belongs to the glycosyl hydrolase family 8. The molecular mass of Cel8A protein induced from Escherichia coli DH5alpha, appeared to be 35 kDa. The optimum pH and optimum temperature was 7.0, and about 30 degrees C for its enzymatic activity respectively.
CONCLUSIONS
R. leguminosarum bv. trifolii 1536 had cel8A gene having an open reading frame of 1047 bp coded for the activity of hydrolyzation of carboxymethyl cellulose.
SIGNIFICANCE AND IMPACT OF THE STUDY
The production of celluloytic enzyme by R. leguminosarum bv. trifolii was confirmed, which would play specific roles in rhizobia. Future study should focus on its role in the infection and nodulation phenomena.
目的
从三叶草根瘤菌1536中分离纤维素酶基因。
方法与结果
采用鸟枪法获得了一个克隆（cel8A），其含有来自三叶草根瘤菌1536的3.1 kb基因组DNA片段。cel8A基因编码348个氨基酸，属于糖基水解酶家族8。从大肠杆菌DH5α诱导表达的Cel8A蛋白的分子量约为35 kDa。其酶活性的最适pH值为7.0，最适温度约为30℃。
结论
三叶草根瘤菌1536具有cel8A基因，其开放阅读框为1047 bp，编码羧甲基纤维素水解活性。
研究的意义和影响
证实了三叶草根瘤菌能产生纤维素酶，该酶在根瘤菌中发挥特定作用。未来的研究应聚焦于其在感染和结瘤现象中的作用。
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