Optimization of buffalo spermatozoa transfection with pbIFN-tau-EGFP gene by simple incubation technique

DOI: 10.5958/0976-0555.2015.00003.5    | Article Id: B-2622 | Page : 14-19
Citation :- Optimization of buffalo spermatozoa transfection with pbIFN-tau-EGFP gene by simple incubation technique.Indian Journal Of Animal Research.2015.(49):14-19
Arvind Kumar, N. Anand Laxmi*, Kuldeep Rai and Ashok Bainda alnand44@gmail.com
Address : DCP Division, National Dairy Research Institute, Karnal-132001, India.

Abstract

The present study was conducted to optimize and analyze the uptake of pbIFN-tau-EGFP by spermatozoa of Murrah buffaloes (Bubalus bubalis) by simple incubation technique. Fresh semen samples from two Murrah buffaloes were selected based on mass activity (+3 and above) from the herd. Three different variables i.e. bovine Interferon tau gene conjugated to enhanced green fluorescent gene ( pbIFN-tau-EGFP) gene concentration, incubation time and number of spermatozoa, were considered for optimization of spermatozoa transfection by simple incubation technique. Before and after transfection, the spermatozoa samples were evaluated for sperm functional parameters, motility and viability. Spermatozoa incubated with different concentrations (10 and 20µg/ml) of  gene (pbIFN tau-EGFP) and without gene which served as control were subjected to  in vitro fertilization (IVF). A significant decrease (P

Keywords

Gene transfer IFN-tau In Vitro fertilization Spermatozoa Transfection.

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