蝗绿僵菌CQMa102体表入侵相关基因MaChy的克隆和分析生物学专业论文.docxVIP

ABSTRACTThe ascomycetes Metarhizium acridum was well characterized entomopathogenic fungi and widely used in biological control programs. The overall host range of M. acridum is broad. During the invasion steps , the formation of appressorium was very important.So cloning and identification of the Characteristics of appressorium formation related gene are necessary to clarify the characteristics of appressorium formation and other important traits related to the molecular mechanism.Based on the Metarhizium acridum CQMa102 subtracted DNA library which constructed by suppression subtractive hybridization (SSH) before penetration.we chose one EST which was highly expressed before penetration. Using the Metarhizium acridum CQMa102 normalized full-length cDNA library during the conidiation stage,we have got the full-length cDNA of MaChy .At the same time,we got the DNA of MaChy through the DNA of Metarhizium acridum CQMa102.In addition, we use RNAi to analyse the function of MaChy gene.① A 1202bp DNA fragment which contained the full-length MaChy gene as wellas upstream and downstream regulatory sequences was cloned and sequenced. The putative coding sequence which contained a 1074 bp ORF, a 80 bp 5’ UTS and 50 bp 3’UTS was identified using ORF finder analysis and BLASTX. A protein of 357 aa with a predicted molecular mass of 39.78kDa and a pI of 5.53 was deduced. Without introns in the full-length DNA , and the deduced protein was predicted with an online program/. (/).② The polypeptide contains four potential site of phosphorylation by caseinkinase 2 (amino acids SELD , THVE , SKHE and STLE at residue 104,134,179 and 341, respectively), four potential site of Protein kinase C phosphorylation (amino acids TIR, SMR, TKK and TPK at residue 4,77,300 and 323, respectively), three potential N-glycosylation sites (amino acids NASK , NASE and NSTD at residues 177,206 and 322, respectively) and four potential myristylation sites (amino acids GTVIN