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Gene Disruption of MNN4 in Candida albican using URA-BLASTER system effects on mannosylphosphrylation

저작시기 2007.01 |등록일 2007.03.04 워드파일MS 워드 (doc) | 15페이지 | 가격 1,700원

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영문 실험 레포트입니다..(조금은 허접합니다.. 참고만 하세요..)
gene disruption using URA-blaster..

목차

INTRODUCTION
MATERIAL AND METHODS
RESULTS
DISCUSSION
REFERENCE

본문내용

The outer cell wall of Candida albicans consists of highly branched glycoproteins modified by N-linked glycosylation including mannosylphosphorylation. Mannosyl-phosphate confers a net negative charge to the cell wall of C. albicans, resulting in binding a positively charged dye Alcian Blue. Previous studies have proposed that mannosylphosphorylation involves in virulence of C. albicans via modification of the outer cell surface. Saccharomyces cerevisiae MNN4 which is required for the mannosylphosphate transfer has high homology with that of C. albicans. The MNN4 gene of C. albicans was disrupted by using Ura-blaster system which was constructed based on the pDH2 plasmid. To select ura- strains, a plate containing 5-fluoroorotic acid was used. After the secondary round transformation, null mutant strains were screened by PCR analysis. The effect of heterozygous mnn4∆ mutants on the cell wall was analysed by Alcian Blue binding assay. The results addressed that homologous mnn4∆ mutant strains showed a significant loss of Alcian Blue binding.

참고 자료

Gow, N.A.R., P.W. Robbing, J.W. Lester, A.J.P. Brown, W.A. Fonzi, T. Chapman and O.S. Kinsman. 1994. A hyphal-secific chitin synthase gene (CHS2) is not essential for growth, dimorphism, or virulence of Candida albicans. Proc. Natl. Acad. Sci. USA 91:6216-6220.

Hobson, R.P., C.A. Munro, S. Bates, D.M. MacCallum, J.E. Cutler, S.E.M. Heinsbroek, G.D. Brown, F.C. Odds, and N.A.R. Gow. 2004. Loss of cell wall mannosylphosphate in Candida albicans does not influence macrophage recognition. The J. of Bio. Chemistry 279(28):39628-39635.

Jigami, Y. and T. Odani. 1999. Mannosylphosphate transfer to yeast mannan. Biochimica et Biophysica Acta 1426:335-345.

Park, Y-N and J. Morschhäuser. 2005. Tetracycline-inducible gene expression and gene deletion in Candida albicans. Eukaryotic Cell 4(8):1328-1342.

Singleton, D.R., J. Masuoka and K.C. Hazen. 2005. Surface hydrophobicity changes of two Candida albicans serotype B mnn4Δ mutants. Eukaryotic Cell 4(4):639-648.

Staab, J.F. and P. Sundstrom. 2003. URA3 as a selectable marker for disruption and virulence assessment of Candida albicans genes. TRENDS in Microbiology 11(2):69-73
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