Using an Improved Site-Directed Mutagenesis for the Creation of Constructs to Analyse Gene Function in Fusarium oxysporum f. sp cubense
文献类型： 外文期刊
作者： Li, C. Y. 1 ; Yi, G. J. 1 ; Chen, S. 1 ; Sun, Q. M. 1 ; Zuo, C. W. 1 ; Huang, B. Z. 1 ; Wei, Y. R. 1 ; Huang, Y. H. 1 ; Wu, 1 ;
作者机构： 1.Guangdong Acad Agr Sci, Fruit Tree Res Inst, Guangzhou, Guangdong, Peoples R China
关键词： S1 nuclease;Site-Directed Mutagenesis;Fusarium oxysporum f. sp cubense (Foc)
期刊名称：INTERNATIONAL ISHS-PROMUSA SYMPOSIUM ON GLOBAL PERSPECTIVES ON ASIAN CHALLENGES
ISSN： 0567-7572
年卷期： 2011 年 897 卷
页码：
收录情况： SCI
摘要： Fusarium oxysporum f. sp. cubense (Foc) is the causal pathogen of Fusarium wilt in banana. Apart from plant resistance, no sustainable solution has been found to control Fusarium wilt. Understanding the genetic basis of pathogenesis in Foc might contribute to developing novel strategies to control Fusarium wilt. We used an improved site-directed mutagenesis to create constructs to analyse gene function in Foc. For the general site-directed mutagenesis, sequencing is the only choice to confirm the mutant clone; however in our protocol, named OE-PCR-S1, we not only improved the efficiency with Dpn I, but also used mismatch cleavage by S1 nuclease to replace sequencing to screen the mutants, which made the protocol more rapid and labour efficient and less expensive. We have used it for Foc, and have successfully deleted three bases from a pathogenic gene, FOW1 (EU795421). The mutant DNA can be inserted into an expression vector and transformed into Foc for functional domain analysis. The overall rate of obtaining the mutant sites was 100%, and the whole mutagenesis process could be completed in less than 2 days, which means that the method is a powerful tool to study pathogenic gene functions and for genetic engineering.
- 相关文献
作者其他论文 更多>>
-
Comparison of volatile aroma compounds in Dwarf Cavendish banana (Musa spp. AAA) grown under organic or traditional cultivation
作者：Dong, T.;Chen, X. J.;Huang, Y. H.;Yi, G. J.;Dong, T.;Chen, X. J.;Huang, Y. H.;Yi, G. J.;Wang, M.
关键词：
-
Core EST-SSR Marker Selection Based on Genetic Linkage Map Construction and Their Application in Genetic Diversity Analysis of Litchi (Litchi chinensis Sonn.) Germplasm Resources
作者：Xiang, X.;Chen, D. M.;Ma, S. P.;Ma, W. C.;Fan, J.;Yang, X. Y.;Liu, M.;Sun, Q. M.
关键词：litchi;EST-SSR;core markers;genetic diversity;hybrids identification
-
The Advancement of Research on Banana Germplasm Resources in China
作者：Wu, Y. L.;Yi, G. J.;Huang, B. Z.;Wei, Y. R.;Li, C. Y.;Hu, C. H.;Huang, Y. H.
关键词：breeding;genetic diversity;Musa spp.;plant genetic resources
-
A study on dietary l-lysine requirement of juvenile yellow catfish Pelteobagrus fulvidraco
作者：Cao, J. M.;Chen, Y.;Zhu, X.;Huang, Y. H.;Zhao, H. X.;Li, G. L.;Lan, H. B.;Chen, B.;Chen, Y.;Pan, Q.
关键词：feed utilization;growth performance;juvenile yellow catfish (Pelteobagrus fulvidraco);l-lysine requirement
-
Construction of cDNA Library and Analysis of SSRs in ESTs of Litchi (Litchi chinensis Sonn.)
作者：Xiang, X.;Sun, Q. M.;Ou, L. X.;Zhao, J. S.;Chen, J. Z.;Cai, C. H.
关键词：litchi;cDNA library;EST;SSR;frequency;characteristics
-
Control of Fusarium wilt in banana with Chinese leek
作者：Huang, Y. H.;Li, C. H.;Zuo, C. W.;Wei, Y. R.;Zhang, L.;Yi, G. J.;Huang, Y. H.;Li, C. H.;Zuo, C. W.;Wei, Y. R.;Zhang, L.;Yi, G. J.;Huang, Y. H.;Wang, R. C.
关键词：Fusarium oxysporum;Panama disease;Banana;Allium tuberosum;Crop rotation;Biocontrol
-
CR-M: an Important Nucleotide Acid Sequence to Distinguish Banana Bunchy Top Virus Isolates from Two Groups
作者：Huang, Y. H.;Zeng, J. W.;Wu, Y. L.;Xia, R.;Yi, G. J.
关键词：
