Cloning, Expression and Activity Assay of Wolbachia DNA Methyl Transferase.

• Main Authors: Yin Hua - Cheng, 鄭尹華
• Other Authors: Wen Yu-Der
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/08280957221687420890

碩士 === 國立彰化師範大學 === 生物學系 === 99 === Wolbachia is one kind of obligated cellular symbiont belongs to alpha Proteobacteria, Rickettsiales and is widely found in insect and arthropod.This intracellular symbiont can cause many reproductive variation of its host,that including male killing, feminization , parthenogenesisand cytoplasmic incompatibility. So far, the causes of these phenomena were not cleard. Wolbachia may be caused methylation by the leafhopper genome., and determined the infected male offspring of the leaf hopper will development female characteristics. This study use the Callosobruchus chinensis and Sitophilus oryzae Linnaeus as model animals and cloned the full-length of Wolbachia DNA methyl transferase in Wolbachia, named Cch_Met and Sli_Met. Furthermore, it is seen the appearance of different mRNA expression of Wolbachia DNA methyltransferase between sexs by RT-PCR ,and the higher expression in female can be observed more than male. Protein expression assay DNA methyltransferase of the pGEM vector, and confirmed the DNA methyltransferase in E. coli BL21 (DE3) by Western blot. Then, methylation of the Wolbachia DNA methyltransferase in E. coli BL21 (DE3) chromosome by isoschizomers (DE3) , and showed methylation in E. coli chromosome.