| Content Provider |
World Health Organization (WHO)-Global Index Medicus |
| Author |
Yang, J. Liu, L. Li, C. Y. Yan, J. L. Wang, H. Yang, Y. Q. Liang, M. L. |
| Description |
Country affiliation: China Author Affiliation: Yang YQ ( Key Laboratory for Agricultural Biodiversity and Pest Management of China Education Ministry, Yunnan Agricultural University, Heilongtan, Kunming, Yunnan, China.); Wang H ( Key Laboratory for Agricultural Biodiversity and Pest Management of China Education Ministry, Yunnan Agricultural University, Heilongtan, Kunming, Yunnan, China.); Liang ML ( Key Laboratory for Agricultural Biodiversity and Pest Management of China Education Ministry, Yunnan Agricultural University, Heilongtan, Kunming, Yunnan, China.); Yan JL ( Key Laboratory for Agricultural Biodiversity and Pest Management of China Education Ministry, Yunnan Agricultural University, Heilongtan, Kunming, Yunnan, China.); Liu L ( Key Laboratory for Agricultural Biodiversity and Pest Management of China Education Ministry, Yunnan Agricultural University, Heilongtan, Kunming, Yunnan, China.); Li CY ( Key Laboratory for Agricultural Biodiversity and Pest Management of China Education Ministry, Yunnan Agricultural University, Heilongtan, Kunming, Yunnan, China.); Yang J ( Key Laboratory for Agricultural Biodiversity and Pest Management of China Education Ministry, Yunnan Agricultural University, Heilongtan, Kunming, Yunnan, China.) |
| Abstract |
The aim of the current study was to investigate the prokaryotic expression of the Magnaporthe oryzae effector genes BAS1 and BAS4 fused to the fluorescent protein mCherry. Based on previous polymorphic analysis of BAS1 and BAS4 in rice blast strains using PCR, blast strains containing the PCR products of BAS1 and BAS4 were selected for liquid culture for total RNA extraction. For PCR analysis, cDNA was selected as a template to amplify the coding region of BAS1 and BAS4, the plasmid pXY201 was selected as template to amplify the mCherry sequence, and the three sequences were cloned into pMD®19-T vectors. Positive recombinant plasmids were digested using two restriction enzymes and the cleaved fragments of BAS1 and mCherry and BAS4 and mCherry were ligated to pGEX-4T-1 vectors and expression was induced using IPTG. The PCR results showed that the sequence sizes of BAS1, BAS4, and mCherry were 348, 309, and 711 bp, respectively, and these were cloned into pMD®19-T vectors. After digestion and gel purification, the fragments of BAS1 and mCherry, BAS4 and mCherry were ligated into pGEX-4T-1 vectors and expressed in Escherichia coli BL21 competent cells. The expressed proteins were approximately 60 kDa, corresponding to their theoretical size. Prokaryotic expression products of BAS1 and BAS4 fused to mCherry were presented in this study, providing a base for constructing prokaryotic expression vectors of pathogen effector genes fused to mCherry, which will contribute to further study of the subcellular localization, function, and protein interactions of these effectors. |
| e-ISSN |
16765680 |
| Journal |
Genetics and Molecular Research |
| Issue Number |
3 |
| Volume Number |
14 |
| Language |
English |
| Publisher |
Fundação de Pesquisas Científicas de Ribeirão Preto |
| Publisher Date |
2015-09-09 |
| Publisher Place |
Brazil |
| Access Restriction |
Open |
| Subject Keyword |
Fungal Proteins Genetics Luminescent Proteins Magnaporthe Recombinant Fusion Proteins Artificial Gene Fusion Cloning, Molecular Dna, Complementary Biosynthesis Chemistry Gene Amplification Gene Expression Genes, Fungal Genetic Vectors Oryza Sativa Microbiology Plasmids Research Support, Non-u.s. Gov't Discipline Genetics Discipline Molecular Biology Discipline Bioinformatics |
| Content Type |
Text |
| Resource Type |
Article |
