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Construction and Expression of an Eukaryotic Expression Vector Containing the Plzf Gene in Spermatogonial Stem Cells of Carp

Author(s): Chinmayee Mohapatra | Hirak Kumar Barman

Journal: Advanced Biotech
ISSN 0973-0109

Volume: 12;
Issue: 11;
Start page: 16;
Date: 2013;
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Keywords: SSCs | Rohu | Plzf | pAcGFP | Promoter

Plzf (Promyelocytic leukemia zinc finger), a transcriptional repressor, is involved in survival and maintenance of pluripotent stem cells including embryonic and spermatogonial stem cells in mammals. Its cDNA was characterized and expression in proliferating spermatogonial stem cells of rohu (Labeo rohita), a farmed carp, was documented. In teleost, the information on its promoter activity is lacking. Here, we have isolated, sequenced and performed the first characterization of regulatory elements for Plzf being expressed in proliferating spermatogonial stem cells of rohu. About 3.2 kb of 5'-flanking region, relative to ATG start codon, derived by Genome Walking was sequenced. Interestingly, computational analysis detected several putative regulatory elements including TATA-box. In this study we described the construction of an eukaryotic expression vector containing Plzf gene and its expression in cultured rohu spermatogonial stem cell. Fluorescence detection displayed that GFP in cytoplasm was high. The recombinant plasmid expression vector was constructed successfully, which may provide a basis for further exploration of function of Plzf in spermatogonial stem cell of fish species
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