Plasmid expressed protein kinase C beta II (PKCbetaII) peptide that selectively inhibits oncogenic VAL 12 ras-p21 induced Xenopus laevis oocyte maturation |
Posted on:2010-10-13 | Degree:M.S | Type:Thesis |
University:Long Island University, The Brooklyn Center | Candidate:Beautreau, Denise N. S | Full Text:PDF |
GTID:2444390002987954 | Subject:Biology |
Abstract/Summary: | |
There are several elements associated with the onset of cancer. Among these is the conversion of a proto-oncogene into the oncogenic product. A single point mutation in the ras-p21 protein allows it to become constitutively active in its GTP bound state, and become an oncogenic product. The oncogenic ras-p21 has been found in 30% of all cancer cases and shows a high incidence specifically in pancreatic carcinomas (>80%).;Protein Kinase C has been shown to be involved in the activation pathway of oncogenic ras-p21. Previous studies conducted by Chie et al. (2003) found the Protein Kinase C Beta-2 (PKCbetaII) peptide corresponding to the V5 catalytic region of the Protein Kinase C Beta-2 gene to inhibit oncogenic ras-p21 induced maturation of Xenopus laevis oocytes. A previous study by Edwards (2006) expressed the PKCbetaII peptide in the phrGFP-II-N (N-) vector to overcome the issue of proteolysis with the peptide experiment of Chie et al. (2003).;This project involved the construction of a plasmid with the DNA sequence encoding the PKCbetaII 645-650 peptide into the pOPRSVI/MCS vector in an attempt to overcome toxic effects caused by the previous vector. The synthesized plasmid was co-injected with Val12 ras-p21 in Xenopus laevis oocytes. The levels of maturation were monitored for a period of 48 hours. The results showed the concentration of 0.3938 mug/muL PKCbetaII plasmid inhibited the oncogenic ras-p21 pathway without having any inhibitory effect on the insulin induced wild-type p21 pathway. The results also showed the enhancement of the inhibitory effect of the PKCbetaII plasmid in the presence of Isopropyl beta-D-1-thiogalactopyranoside (IPTG). |
Keywords/Search Tags: | Pkcbetaii, Plasmid, Protein kinase, Oncogenic, Xenopus laevis, Ras-p21, Peptide, Induced |
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