In order to obtain the appropriate CHO (Chinese Hamster Ovary) cell line overexpressing 5-hydroxytryptamine 1A receptor gene (HTR1A gene, human cDNA clone ref. ID NM_000524) pcDNA 3.1 (+) and pCMV6-XL4/HTR1A plasmids  were processed with Not I restrictive enzyme and ligated.  The obtained pcDNA 3.1(+) vector containing HTR1A gene was propagated in competent E. coli cells on LB-agar plates containing ampicilin. Individual clones containing the pcDNA/HTR1A plasmid were picked up and cultured in the LB Broth medium.

After isolation the pcDNA/HTR1A plasmid has been transferred into CHO-K1 (clone-1)cells with the aid of FuGene reagent. Then the cells were cultured in the presence of geniticine. Selected single cells were cloned and the expression of pcDNA/HTR1A plasmid was evaluated with the aid of Western blot analysis.

The resulting clone CHO-K1 cell lines with stable overexpression of the gene HTRA1 were used as a model for testing the functional activity of 5HT_1A receptor ligands.

Acknowledgements

This study was supported by a grant PNRF-103-AI-1/07 from Norway through the Norwegian Financial Mechanism.

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