Human cyclin A & B toxic to yeast?

Zhonglin Chai zhonglin.chai at med.monash.edu.au
Wed Jul 19 19:07:48 EST 2000


Hello Everyone,

I am trying to test the interaction of my protein of interest with human
cyclin A and B using yeast 2-hybrid system. I am not able to generate
any yeast transformants with human cyclin constructs in either pLex A
(BD) or pGAD 10 vector (AD), while human cdc2 kinase (in both vectors)
and all the other constructs of my gene of interest are successfully
transformed. Lew, et al (1991) (Cell 66:1197) showed that "gross
overexpression" of human cyclin B2 is toxic to yeast cells and human
cyclin B2 toxic when the yeast cells express its own cyclin CLN2,
although they, in lower expression levels, were used to resuce the
genetic defect of yeast G1 cyclins. If this is the case I wonder if
there is a yeast 2-hybrid system that allow low expression levels to
test protein-protein interactions?  Do you know human cyclin A or B has
been previously used in yeast 2-hybrid system? Do you know any other way
to get around the problem?

Regards.

ZhongLin Chai, PhD
__________________________________________________________
Department of Pathology and Immunology
Monash University Medical School
Alfred Hospital
Commercial Rd, Prahran, VIC 3181, AUSTRALIA
Telephone: (61 3) 9903 0698 (lab)
           (61 3) 9903 0696 (office)
Mobile:    0413 58 1940 or International: +61 413 58 1940
Fax:       (61 3) 9903 0731
email:     zhonglin.chai at med.monash.edu.au
___________________________________________________________







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