Malaysian Abstracting and Indexing SystemIntegration of the Toxoplasma gondii MIC2 Gene Into the Pichia pastoris Genome Yee, Wai-Yanauthor Wah, Kiew-LianauthorThe Toxoplasma gondii MIC2 gene encodes a microneme protein that is believed to play an important role in the adhesion of the parasite to its host cell. Since the Pichia pastoris expression system is capable of producing biologically active foreign proteins, expression of recombinant MIC2 in this yeast will potentially provide useful MIC2 material for functional studies of this protein. As a prior effort to utilise this eukaryotic expression system in the study of MIC2, we have integrated the MIC2 gene into the genome of P. pastoris. Amplification of the MIC2 gene was carried out using the polymerase chain reaction (PCR) technique and cloned into the pCR2.1 vector. The resulting pCR2.1-MIC2 recombinant plasmid was transformed into the Escherichia coli TOP 10 strain. Positive colonies were determined by DNA plasmid analysis. The pCR2.1-MIC2 recombinant plasmid was isolated, and digested with EcoRI and XbaI restriction enzymes. The MIC2 fragment was then purified and cloned into the pPICZ B expression vector. The resulting pPICZ-MIC2 recombinant plasmid was transformed into the E. coli DH5α strain. Positive transformants were determined by PCR analysis. The pPICZ-MIC2 recombinant plasmid was extracted, linearised and transformed into P. pastoris via electroporation. The integration of MIC2 into the P. pastoris genome was subsequently demonstrated by PCR.Q Science, Computer ScienceR Medicine, Dentistry, Pharmacy, Nursing2005Malaysian Society for Biochemistry and Molecular BiologyJournal

For work being deposited by its own author: In self-archiving this collection of files and associated bibliographic metadata, I grant Malaysian Abstracting and Indexing System the right to store them and to make them permanently available publicly for free on-line. I declare that this material is my own intellectual property and I understand that Malaysian Abstracting and Indexing System does not assume any responsibility if there is any breach of copyright in distributing these files or metadata. (All authors are urged to prominently assert their copyright on the title page of their work.)

For work being deposited by someone other than its author: I hereby declare that the collection of files and associated bibliographic metadata that I am archiving at Malaysian Abstracting and Indexing System) is in the public domain. If this is not the case, I accept full responsibility for any breach of copyright that distributing these files or metadata may entail.

Clicking on the deposit button indicates your agreement to these terms.