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Corn leaf blight caused by Exserohilum turcicum maize leaf disease prevention corn leaf blight is the most economical and effective way to nurture and grow corn resistant varieties. Therefore, the separation Corn Resistance to clone genes, a clear identity and function of the encoded protein, has an important significance for revealing the Corn Resistance molecular mechanisms and resistance gene engineering breeding. The test turcica B37Htl with corn 0 Souchong and the 1st race incompatible combination and affinity combinations as test material, the use of mRNA differential display technology (DDRT-PCR) of corn leaves corn inbred lines gene expression analysis, recovery of non-affinity interaction specifically expressed gene fragments, cloning, sequencing, and homology analysis. Test to obtain the following main results: (1) were amplified by PCR and polyacrylamide gel electrophoresis, screening 26 random primers. It was found that six primers for step amplification method, the annealing temperature in the range of 40 to 47 ° C; 9 primer suitable for a two-step amplification method, the annealing temperature (second step amplification) in between 46 to 51 ° C. ② DDRT-PCR amplification of cDNA sample using random primers screened by polyacrylamide gel electrophoresis and silver staining, a total of 107 differentially expressed genes. These fragments can be divided into three categories: A. affinity interaction and control non-affinity interactions in specific fragments of 26, accounting for 24%; B. and affinity interaction and compared to the control, non-affinity interaction the fragments expressed increased to 39 (36%); C. affinity interaction and compared to the control, non-affinity interaction in decreased expression fragment 42, accounting for 40% . Their recovery and secondary amplification detected by 1.2% agarose gel electrophoresis, wherein 98 of the recovered product to produce the amplified bands, differences strip recovery was 92%. ③ After the trans Northern blot analysis, and ultimately positive fragment 46, the positive rate was 46.9%, including 10 Class A fragment. Accounted for 21.8%: The B class fragment 19, accounting for 41.1%; C class fragments of 17, accounting for 37.1%. ④ elect five Class A fragment cloning, sequencing and homology analysis, and found that they are lower than known protein sequence homology, 30% to 40%, the new gene. Further access to the full length cDNA and by functional complementation experiments to determine the functionality needed for these fragments.
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