%0 Journal Article
%T Cloning of β-1,3-glucanase gene from Trichoderma virid LTR-2 and its expression in Pichia pastoris<br>绿色木霉LTR-2 β-1, 3-葡聚糖酶基因的克隆及其在 毕赤酵母中的表达
%A Wen Gao
%A Yuanzheng Wu
%A Hetong Yang
%A <br>高雯
%A 吴远征
%A 杨合同
%J 微生物学报
%D 2008
%I 
%X Text We designed a pair of primers according to fungal glucanase genes obtained from GenBank and cloned a novel beta-1,3-glucanase gene (glu) from Trichoderma virid LTR-2 cDNA by PCR. Then we linked the fragment with pMD18-T vector and sequenced it. The sequence analysis indicated that glu was composed of 2289 nucleotide residues. The fragment contained an Open Reading Frame coding 762 amino acids and was similar to previous reports. Translated amino acids sequence of glu contained two Conservative Districts of beta-1,3-glucanase which were RVVYIPPGTY and AASQNKVAYF. By nucleotide blasting in NCBI glu showed high homology to three beta-1,3-glucanase genes from Trichoderma sp., especially with T.harzianum bgn3.1 and Hypocrea virens bgn13.1, which the homology reached 93%. The sequence was submitted to GenBank and the Accession Number is EF176582. Then we connected glu gene with the Pichia pastoris shuttle vector-pPIC9K. The recombinant plasmid named pGLU14 was transformed into methylotropic yeast P. pastoris KM71 after linearization. The recombinant strain KGLU14 expressing beta-1,3-glucanase at high level was obtained through plate screening. The SDS-PAGE result indicated that molecular weight of the recombinant beta-1,3-glucanase was about 80kDa and the activity of the recombinant enzyme could reach 889U/mL in liquid culture.
%K β-1
%K 3-葡聚糖酶
%K 克隆
%K 序列分析
%K 分泌表达
%K 绿色
%K 木霉
%K 葡聚糖酶基因
%K 克隆
%K 毕赤酵母菌株
%K 分泌表达
%K Cloning
%K Pichia
%K pastoris
%K expression
%K Trichoderma
%K 培养基
%K 摇瓶发酵
%K 测值
%K 理论
%K 分子量
%K 结果
%K 电泳
%K 酵母基因组
%K 整合
%K 菌落
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=24289A81F89D0509381A220A0ACC6F8A&yid=67289AFF6305E306&vid=B6DA1AC076E37400&iid=0B39A22176CE99FB&sid=1DF3F9D75A12D97B&eid=B78CD622C1934236&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=12