ISSN 2756-3707
International Journal of Histology and Cytology ISSN 2447-9535 Vol. 5 (1), pp. 375-380, January, 2018. © International Scholars Journals
Full Length Research Paper
A study of how a Chinese Hamster Ovary cell line can translate a functional recombinant Dsn1p from the yeast
Beow Chin Yiap1, A .K. Radhakrishnan2 and Nadimpalli Ravi S. Varma3*.
1School of Pharmacy and Health Sciences, Faculty of Medicine and Health, International Medical University, No. 126, Jalan 19/155B, Bukit Jalil, 57000 Kuala Lumpur, Malaysia.
2Department of Pathology, Faculty of Medicine and Health, International Medical University, No. 126, Jalan 19/155B, Bukit Jalil, 57000 Kuala Lumpur, Malaysia.
3Department of Biological and Agriculture Engineering, University of California, Davis, 95616, California, USA.
*Corresponding author. E-mail: [email protected]. Tel: 001-530-754-0332. Fax: 530 752-2640.
Accepted 11 April, 2017
Abstract
Dsn1p is a component of the mind complex (Mtw1p Including Nnf1p-Nsl1p-Dsn1p) that is essential for the segregation of chromosome in yeast cells. This protein assists the joining of kinetochore subunits contacting DNA to those contacting microtubules in yeast cells. Null mutants of this protein are not viable while the over-expression of the Dsn1 protein in yeast cells results in nuclear anomalies and growth defects. In this paper, we show that tetrad analysis indicates haplo-insufficiency and segregational errors in yeast diploid single deletants. Expression of Dsn1p in CHO has been achieved using the pcDNA 3.1/HIS A expression vector. Analysis by DNA sequencing showed no changes in the DSN1 DNA sequence. We also showed that nuclear fragmentation and cellular morphological changes takes place in CHO cell line that were successfully transfected with the pcDNA 3.1/HIS A expression vector containing the Dsn1p.
Key words: Saccharomyces cerevisiae, kinetochore, Dsn1p, Chinese Hamster Ovary (CHO).