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  • [Screening and identifying cellulose degrading bacteria associated with Bursaphelenchus xylophilus and cloning corresponding genes].

[Screening and identifying cellulose degrading bacteria associated with Bursaphelenchus xylophilus and cloning corresponding genes].

Wei sheng wu xue bao = Acta microbiologica Sinica (2013-02-07)
Qiuhong Niu, Lin Zhang, Xueying Chu, Fengguang Du, Wensheng Feng, Fengli Hui, Tao Ke, Yongxin Fan
ABSTRACT

To screen, identify bacterial strains with high capability to degrade cellulose from bacteria associated with Bursaphelenchus xylophilus and to clone related genes. First, we collected B. xylophilus samples from pine wood nematode disease areas in Nanyang, Henan province, China. Then, we obtained the bacterial strains with high cellulase activities by primarily screening according to Congo red plate methods. The bacterial strain was classified by phenotypic and genotypic characteristics. We designed degenerate primes according to the known endoglucanase gene sequences in GenBank to carry out PCR, and analyzed the cloned sequence. We obtained seven bacterial strains with high cellulase activities. Among them, the bacterial strain numbered C8 showed the highest cellulase activities. The bacterium was classified to be Enterobacter genus. The full length of a cellulase gene cDNA (1104 bp) (GenBank JQ845065) coding region was successfully cloned. The homogeneous analysis demonstrated that the deduced nucleotide and amino acid of the gene separately shared 97% and 92% with the cellulase from E. aerogenes KCTC 2190, and 82% with the endo-1,4-D-glucanase gene from Klebsiella pneumoniae, and 82% with the a cellulase gene from unculturable bacteria. It was a novel cellulose gene cloned from B. xylophilus associated bacteria.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Cellulase from Aspergillus niger, powder, ≥0.3 units/mg solid
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Cellulase from Trichoderma reesei ATCC 26921, lyophilized powder, ≥1 unit/mg solid
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Sigma-Aldrich
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Sigma-Aldrich
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