Detection of phospholipase C zeta in testis of Rattus argentiventer Robinson & Kloss (rice-field rat) using molecular techniques
Phospholipase C-zeta (PLC-ζ) is a specific enzyme found in sperm of mammals responsible for triggering calcium oscillations leading to oocyte activation during fertilization. The activation causes the fertilized oocyte to divide and develop into an embryo. Rattus argentiventer (Rice Field Rat) is of...
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| Format: | Thesis |
| Language: | English |
| Published: |
2013
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| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/71007/1/FPSK%28M%29%202017%2019%20IR.pdf |
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| Summary: | Phospholipase C-zeta (PLC-ζ) is a specific enzyme found in sperm of mammals responsible for triggering calcium oscillations leading to oocyte activation during fertilization. The activation causes the fertilized oocyte to divide and develop into an embryo. Rattus argentiventer (Rice Field Rat) is often responsible for destruction of agricultural crops specifically paddy fields. The mammal is known for its rapid reproductive potential and as a carrier of pathogen. This rodent pest population is currently being controlled with baits, traps and biological control such as Tyto alba. However the current methods are quite expensive, needs long term monitoring and are hazardous to the environment. Thus the knowledge of fertility such as sperm factor of this rodent could be helpful in a way to prevent overpopulation of this rodent. Two detection methods to identify PLC-ζ gene fragments from the testis of Rattus argentiventer using conventional Polymerase Chain Reaction (Reverse Transcriptase RT-PCR) and Real Time Polymerase Chain Reaction (qRT-PCR) techniques were used in order to obtain the bands of PLC-ζ. Following that, sequencing of DNA structure and cloning of PLCwere performed to amplify gene fragment of PLC-ζ. Approximately 420 bp nucleotides identified sequence was then keyed in into the Basic Local Alignment Search Tool (BLAST) portal in National Centre of Bioinformatics Information (NCBI) for comparison with standard nucleotide sequences from other species for sequence alignment. The specific target 420 bp nucleotides of PLC-ζ were cloned using a kit through ligation and transformation. The result showed that PLC-ζ enzyme was present in Rattus argentiventer using PCR technique. However, the cloning procedure carried out was not able to show the presence of target gene fragments of the enzyme. It is confirmed that PLC-ζ enzyme was present in Rattus argentiventer through the two detection methods used. The study is a novel study to detect of PLC-ζ in paddy rats using molecular approaches. The result obtained with RT-PCR was validated with qRT-PCR and the sequence obtained showed excellent similarity homology with published sequence of PLC-ζ in Rattus norvegicus. As such this would give valuable baseline information for future researches to be carried out in the approach of controlling and preventing the continous growth of Rattus argentiventer population. |
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