Construction of a yeast one-hybrid cDNA library and CcCas5 promoter bait vector of Corynespora cassiicola causing leaf fall disease of Hevea brasiliensis
In order to screen the transcription factors that bind to the CcCas5 promoter,a yeast one-hybrid cDNA expression library of Corynespora cassiicola and a CcCas5 promoter bait vector were constructed,and the optimal inhibitory concentration of 3-amino-1,2,4-triazole(3-AT)was screened in this study.The results showed that the total capacity of the constructed yeast one-hybrid cDNA expression library was 3.49 × 106 cfu,the inserted fragments mainly distributed between 750 to 2 000 bp,and the recombination rate was 95.8％.The 2 600 bp promoter sequence of CcCas5 was cloned and the binding sites of 165 main fungal transcription factors in 10 families were predicted.Yeast one-hybrid promoter bait vectors carrying 1 500 bp and 1 000 bp promoter sequences were constructed,respectively,and 10 mmol/L of 3-AT could inhibit the background expression of pHis2.1-pCcCas5-1000 and pHis2.1-pCcCas5-1500.The above results lay the foundation for screening transcription factors that bind to the CcCas5 promoter through yeast one-hybrid screening and further studying the regulatory effects of transcription factors on CcCas5 expression.
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