Construction of Promoter Probe Vectors,and Screening and Identification of Oidium heveae Promoters

doi:10.13560/j.cnki.biotech.bull.1985.2018-0584

Abstract

Abstract: This study aims to construct a probe vector suitable for rapidly detecting the promoter of Oidium heveae HO-73. The kanamycin-resistance gene was used as a reporter gene to construct the promoter probe vector pUC19-K in the pUC19 backbone vector,and it was ligated with the CaMV35S promoter to verify the function of the promoter probe vector. The promoter probe vector pUC19-K was employed to screen and identify the predicted promoter fragments LY1,LY2,LY3,and LY4. The CaMV35S promoter was ligated into the promoter probe vector to obtain the promoter probe vector pUC19-K that can detect the promoter activity. Using bioinformatics software to predict the whole genome data of Oidium heveae HO-73,4 theoretically active promoter sequences,LY1,LY2,LY3,and LY4,were obtained,and their activities were compared using the constructed promoter activity probe vector. The Kana tolerance test showed that strains containing LY2 and LY3 were more tolerant with increasing concentrations of kanamycin,resulting in two more active promoters,LY2 and LY3.The above results indicate that the constructed promoter activity probe vector can be effectively and sensitively used for screening the strong promoter of HO-73 and detecting promoter activity.

Key words: Oidium heveae, promoter, probe vector

XU Liang-xiang, LIU Yao, LIAO Xiao-miao, WANG Yi, M.J.N. RAJAOFERA, HE Qi-guang, LIU Wen-bo, LIN Chun-hua, MIAO Wei-guo. Construction of Promoter Probe Vectors,and Screening and Identification of Oidium heveae Promoters[J]. Biotechnology Bulletin, 2019, 35(1): 51-57.

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