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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Main Author:	Amir Shapuddin, Fadzlina
Format:	Thesis
Language:	English
Published:	2013
Subjects:	Fertilization - genetics Sperm-Ovum Interactions Spermatozoa
Online Access:	http://psasir.upm.edu.my/id/eprint/71007/1/FPSK%28M%29%202017%2019%20IR.pdf
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spelling	my-upm-ir.710072019-09-20T02:48:33Z Detection of phospholipase C zeta in testis of Rattus argentiventer Robinson & Kloss (rice-field rat) using molecular techniques 2013-05 Amir Shapuddin, Fadzlina 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. Fertilization - genetics Sperm-Ovum Interactions Spermatozoa 2013-05 Thesis http://psasir.upm.edu.my/id/eprint/71007/ http://psasir.upm.edu.my/id/eprint/71007/1/FPSK%28M%29%202017%2019%20IR.pdf text en public masters Universiti Putra Malaysia Fertilization - genetics Sperm-Ovum Interactions Spermatozoa
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
topic	Fertilization - genetics Sperm-Ovum Interactions Spermatozoa
spellingShingle	Fertilization - genetics Sperm-Ovum Interactions Spermatozoa Amir Shapuddin, Fadzlina Detection of phospholipase C zeta in testis of Rattus argentiventer Robinson & Kloss (rice-field rat) using molecular techniques
description	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.
format	Thesis
qualification_level	Master's degree
author	Amir Shapuddin, Fadzlina
author_facet	Amir Shapuddin, Fadzlina
author_sort	Amir Shapuddin, Fadzlina
title	Detection of phospholipase C zeta in testis of Rattus argentiventer Robinson & Kloss (rice-field rat) using molecular techniques
title_short	Detection of phospholipase C zeta in testis of Rattus argentiventer Robinson & Kloss (rice-field rat) using molecular techniques
title_full	Detection of phospholipase C zeta in testis of Rattus argentiventer Robinson & Kloss (rice-field rat) using molecular techniques
title_fullStr	Detection of phospholipase C zeta in testis of Rattus argentiventer Robinson & Kloss (rice-field rat) using molecular techniques
title_full_unstemmed	Detection of phospholipase C zeta in testis of Rattus argentiventer Robinson & Kloss (rice-field rat) using molecular techniques
title_sort	detection of phospholipase c zeta in testis of rattus argentiventer robinson & kloss (rice-field rat) using molecular techniques
granting_institution	Universiti Putra Malaysia
publishDate	2013
url	http://psasir.upm.edu.my/id/eprint/71007/1/FPSK%28M%29%202017%2019%20IR.pdf
_version_	1747812952798396416

Detection of phospholipase C zeta in testis of Rattus argentiventer Robinson & Kloss (rice-field rat) using molecular techniques

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