The life cycle of Giardia lamblia contains two differentiation processes, encystation and excystation. We performed an experiment to identify the genes induced during encystation using the differential display reverse transcriptase-polymerase chain reaction. Three of twelve isolated cDNA clones that showed increased transcription during encystation were identified to be of the myb2, which encodes a well-known transcriptional factor involved in cellular development and differentiation. The amino acid sequences of the Myb2 protein deduced from the isolated gene revealed that this Myb2 has a DNA binding domain comprising two imperfect repeats at its carboxyl-terminus. The nuclear localization of Myb2 protein during encystation was observed in vivo by expressing a Myb2-GFP fusion protein. In a random site selection experiment, the oligonucleotides bound by rMyb2 contained a conserved sequence of GTTT(G/T)(G/T). Two promoters of the encystation-induced genes, myb2, and cwp1, were also found to bind to rMyb2, whereas gap1, one of the constitutive genes did not.
Bibliographical noteFunding Information:
This work was supported by a Korean Research Foundation Grant (KRF2001-015-FP0079). We thank Prof. Steve Singer of Georgetown University for providing a plasmid pGFP.pac and the transfection protocol, and Prof. Kyu-Ho Lee of Hankuk University of Foreign Studies for critical comments on this manuscript.
All Science Journal Classification (ASJC) codes
- Molecular Biology