Isolation and detection of PLC (Sperm Factor Protein) in testes of Rattus rattus diardii using one-step RT-PCR


Citation

Lee L.H., . and Fadzlina A.S., . and Cheah Y.K., . and Nasyriq A.M.N., . and Noor Faezah M.N., . and Sukardi S., . Isolation and detection of PLC (Sperm Factor Protein) in testes of Rattus rattus diardii using one-step RT-PCR. pp. 47-52. ISSN 1394-3227

Abstract

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.


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Abstract

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.

Additional Metadata

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Item Type: Article
AGROVOC Term: Rats
AGROVOC Term: Rattus
AGROVOC Term: Fertilization
AGROVOC Term: Reproductive physiology
AGROVOC Term: Overpopulation
AGROVOC Term: Reproduction control
AGROVOC Term: Contraception in animals
AGROVOC Term: Biological control
Depositing User: Ms. Suzila Mohamad Kasim
Last Modified: 24 Apr 2025 06:27
URI: http://webagris.upm.edu.my/id/eprint/21541

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