A study of how a Chinese Hamster Ovary cell line can translate a functional recombinant Dsn1p from the yeast

Abstract


Beow Chin Yiap, A .K. Radhakrishnan and Nadimpalli Ravi S. Varma*

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.

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