中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > β-葡萄糖苷酶bglⅠ基因在粟酒裂殖酵母中的表达及功能分析 西北农林科技大学学报(自然科学版) 2012,40 (12) 171-176
Position: Home > Articles > Expression and functional analysis of a β-glucosidase gene in Schizosaccharomyces pombe Journal of Northwest A & F University(Natural Science Edition) 2012,40 (12) 171-176

β-葡萄糖苷酶bglⅠ基因在粟酒裂殖酵母中的表达及功能分析

作  者：
付永平;李博;王丕武;高玮;夏海丰;张卓
单  位：
通化市农业科学院;吉林农业大学农学院;吉林市农业科学院
关键词：
绿色木霉;β-葡萄糖苷酶;粟酒裂殖酵母;酶学活性
摘  要：
【目的】克隆绿色木霉β-葡萄糖苷酶bglⅠ基因,并在粟酒裂殖酵母细胞中进行表达分析。【方法】利用PCR技术,从绿色木霉中克隆纤维素酶β-葡萄糖苷酶bglⅠ基因的DNA序列,再通过XhoⅠ和NotⅠ2个酶切位点与粟酒裂殖酵母表达载体pREP5N连接,构建酵母真核表达载体,利用电转化方法转化粟酒裂殖酵母SP-Q01细胞,并在SP-Q01中进行表达分析,最后采用DNS法测定表达产物的酶学活性。【结果】PCR扩增得到2 380bp的bglⅠ基因序列,包括完整的CDS序列和信号肽序列,编码序列与已报道的里氏木霉纤维素酶基因序列相似性为100%;对酵母转化子进行诱导培养后,SDS-PAGE电泳检测得到大小为76ku的目的蛋白条带,在SP-Q01细胞中重组酶活性最高达155U/mL,最佳培养时间为72h,产生酶活的最适温度为60℃。【结论】成功克隆出了绿色木霉β-葡萄糖苷酶bglⅠ基因的DNA序列,该基因能在粟酒裂殖酵母细胞中成功表达,为绿色木霉β-葡萄糖苷酶bglⅠ基因的深入研究和应用奠定了基础。
译  名：
Expression and functional analysis of a β-glucosidase gene in Schizosaccharomyces pombe
作  者：
FU Yong-ping1,LI Bo2,WANG Pi-wu1,GAO Wei3,XIA Hai-feng1,3,ZHANG Zhuo1(1 College of Agriculture,Jilin Agricultural University,Changchun,Jilin 130118,China;2 Jilin City Academy of Agricultural Sciences,Jilin,Jilin 132101,China;3 Tonghua City Academy of Agricultural Sciences,Tonghua,Jilin 135000,China)
关键词：
Trichoderma viride;bglⅠ gene;Schizosaccharomyces pombe;cellubiohydrolase activity
摘  要：
【Objective】 The study aimed to clone β-glucosidase(bglⅠ) gene from Trichoderma viride and analyze its expression in Schizosaccharomyces pombe.【Method】 The bglⅠ gene was cloned by PCR method,and inserted into the expressing vector pREP5N of Schizosaccharomyces pombe to generate a recombinant plasmid pREP5N-bglⅠ.The recombinant pREP5N-bglⅠ plasmid was transformed into S.pombe SP-Q01 by electroporation method.Using cellubiohydrolase activity test and SDS-PAGE analysis,the bglⅠ was expressed by S.pombe-bglⅠ.【Result】 Sequence analysis showed that PCR amplification generated a gene sequence with 2 380 bp,including coding sequence and signal peptide sequence.The DNA sequence and the putative amino acid sequence shared 100% identity with the reported sequence.The molecular weight of purified protein product from S.pombe-bglⅠ was 76 ku.The cellubiohydrolase activity of the cultivation supernatant of recombinant S.pombe-bglⅠ was 155 U/mL.The enzyme activity culminated when being cultured for 72 h,and the optimal enzyme reaction temperature was 60 ℃.【Conclusion】 The bglⅠ gene from Trichoderma viride was cloned,and its function expression in eukaryotic cells was analyzed.This could provide a foundation for further research and application of the cellulase bglⅠ gene from Trichoderma viride.

相似文章

导出引用
计量
文章访问数: 5
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46