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Manuscript Details

Isolation and Detection of PLC? (Sperm Factor Protein) in Testes of Rattus rattus diardii Using One-Step RT-PCR

Nasyriq, A.M.N., Fadzlina, A.S., Noor Faezah, M.N., Lee, L.H., Cheah Y.K. and Sukardi*, S.

Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia.

Malaysian J. Anim. Sci. 2013 16(1): 47-52

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The fertilization process in many species studied involves induction of Ca2+ oscillations in eggs. Prolonged time period of calcium oscillations is essential for regulating various kinds of events that will occur after fertilization. PLC zeta (PLC?) has been discovered as a novel isoenzyme of PLC which is responsible in promoting intracellular Ca2+ oscillation in oocytes. Until now, PLC? has been identified in many mammalian species. Although PLC? has been identified in the rodent family, such as the laboratory mice, rats and hamsters, no research has been carried out to identify PLC? in house rats. This study was carried out in order to identify the existence of PLC? in testes of house rats (Rattus rattus Diardii), one of the major pests in houses. Testes from ten male rats were subjected to RNA extraction using easy-BLUETM Total RNA Extraction Kit. One-step RT-PCR technique was chosen to identify PLC? using forward and reverse primers from published sequence of PLC-zeta. PCR amplicons were extracted from Agarose gel, purified and further subjected to sequencing. Sequences obtained were aligned using NCBI Blast search at www.ncbi.nlm.nih.gov/blast to check for identification from published PLC-zeta sequences. Results from this study showed that PLC? from the testes of house rat was successfully amplified by using One-step RT-PCR technique. The sequence also showed strong homology to published sequence of Rattus norvegicus PLC?. We hope that with the understanding of molecular properties of PLC? in house rats could lead to development of contraceptive vaccines, an alternative approach to control the rat population.