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| Technique for Exploring Pectinase Gene of Bast Fiber Degumming from Enrichment Liquid |
| DUAN Sheng-wen, LIU Zheng-chu, ZHENG Ke, FENG Xiang-yuan, CHENG Li-feng, ZHENG Xia |
| Instititute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha, Hunan 410205, China |
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Abstract To break through the deficiency of traditional culture technique in screening pectinase gene for bast fiber bio-degumming, different methods of total DNA extraction, sample enrichment, and optimization of PCR reaction system were designed, and a set of metagenomic technique that was suitable for excavation of pectinase gene for bast fiber degumming was established. The results showed that, degumming time of shake cultivation was shorter 51.5% than that of static cultivation to enrich microorganisms of ramie degumming; the total DNA for the study of metagenomic technique was extracted from the fourth fermentation by the PowerSoil? DNA Isolation Kit; the PCR optimal reaction system in the total volume of 25 ml: the concentrations of Mg2+, dNTPs, primer, DNA template were 2.0 mmol/L, 0.2 ng/μl, 0.6 μmol/l, 2.5 ng/μL, and 0.8 U respectively. The gene sequence obtained by the optimized technique was completely similar to the pectinase gene FJ538208 sequence reported.
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Received: 30 October 2013
Published: 25 January 2014
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[1] Fu J J, Li X Q, Gao W D, et al. Bio-processing of bamboo fibres for textile applications: a mini review. Biocatal Biotransform, 2012, 30: 141-153.
[2] 刘正初. 生物制剂在草本纤维质农产品加工业中的应用进展. 中国农业科技导报, 2013, 15(5): 17-23. Liu Z C. Progress of biological agents used in process industries for agricultural products of herbaceous fibers. Journal of Agricultural Science and Techology, 2013, 15(5): 17-23.
[3] Brühlmann F, Keen N T. Cloning, sequencing and expression of the pel gene from an Amycolata sp. Gene, 1997, 202: 45-51.
[4] 李琦, 戴小阳, 刘正初, 等. 欧文氏菌T85-166果胶酶基因克隆与序列分析. 湖南农业科学, 2009, (7): 8-10. Li Qi, Dai X Y, Liu Z C, et al. Gene Cloning and sequence analysis on pectinase of Erwinia T85-166. Hunan Agricultural Sciences, 2009, (7): 8-10.
[5] Singh R, Dhawan S, Singh K, et al. Cloning, expression and characterization of a metagenome derived thermoactive/thermostable pectinase. Mol Biol Rep, 2012, 39: 8353-8361.
[6] 苑鹏. 果胶酶基因的克隆表达及其在小尾寒羊瘤胃环境中的多样性分析. 北京: 中国农业科学院, 2012. Yuan P. Gene Cloning and Expression of Pectinases and Their Diversity Analysis in the Rumen of Small Tail Han Sheep. Beijing: China Academy of Agricultural Sciences, 2012.
[7] 王德良, 周小慧. 基础生物学实验教程. 北京: 中国科学技术出版社, 2006. 447-449. Wang D L, Zhou X H. Basic Biology Experiment Course. Beijing: China Science and Technology Press, 2006. 447-449.
[8] 黄循柳, 黄仕杰, 郭丽琼, 等. 宏基因组学研究进展. 微生物学通报, 2009, 36(7): 1058-1066. Huang X L, Huang S J, Guo L Q, et al. Advances of metagenomics. Microbiology, 2009, 36(7): 1058-1066.
[9] 孙欣, 高莹, 杨云锋. 环境微生物的宏基因组学研究新进展生物多样性. 2013, 21 (4): 1-8. Sun X, Gao Y, Yang Y F. Recent advancement in environmental research with metagenomics tools. Biodiversity Science, 2013, 21(4): 1-8. |
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