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Published on: August 29, 2020
Chromosomal mapping, sequence and transcription analysis of the porcine fertilin beta gene (ADAM2)
A E Day1, C R Quilter, C A Sargent
1Sygen International Group, Department of Pathology, University of Cambridge, Cambridge, UK. aed21@cam.ac.uk
Abstract:
Fertilin beta (ADAM2) forms a part of the heterodimeric surface protein fertilin, found on the plasma membrane of mammalian sperm, and has been implicated in the process of sperm-egg fusion. Analysis of cDNA products obtained from adult porcine testis mRNA has presented a sequence corresponding to 2620 bp of the ADAM2 gene. This sequence contained an open reading frame encoding a 735-amino acid protein and homologous to ADAM2 genes known in other mammalian species. Polymerase chain reaction (PCR) analysis of genomic DNA showed that the 2620 bp of cDNA sequence comprises at least 21 exons and spans approximately 76 kb of genomic DNA, with its size and structure being relatively conserved between mouse, human and pig. Fluorescence in situ hybridization was used to map ADAM2 to chromosome 15 of the pig, using a bacterial artificial chromosome clone from the PigE BAC library. This finding is consistent with comparative mapping experiments performed between pig and human chromosomes. Analysis of nine mRNA samples, by reverse transcriptase-PCR, from different porcine tissues has also suggested that expression of ADAM2 is limited to the testis, a finding that is consistent with other mammalian species.
Insights
Fertilin beta (ADAM2) is crucial for sperm-egg fusion. This study details the ADAM2 gene
Area of Science:
- Reproductive Biology
- Genetics
- Molecular Biology
Background:
- Fertilin beta (ADAM2) is a key protein in mammalian sperm involved in sperm-egg fusion.
- Understanding ADAM2's genetic structure and expression is vital for reproductive research.
Purpose of the Study:
- To characterize the porcine ADAM2 gene sequence and structure.
- To determine the chromosomal location of ADAM2 in pigs.
- To investigate the tissue-specific expression of ADAM2 in pigs.
Main Methods:
- cDNA sequencing of porcine testis mRNA.
- Polymerase Chain Reaction (PCR) for genomic DNA analysis.
- Fluorescence In Situ Hybridization (FISH) for gene mapping.
- Reverse Transcriptase-PCR (RT-PCR) for mRNA expression analysis.
Main Results:
- A 2620 bp cDNA sequence for porcine ADAM2 was identified, encoding a 735-amino acid protein.
- The ADAM2 gene comprises at least 21 exons spanning approximately 76 kb of genomic DNA.
- ADAM2 was mapped to pig chromosome 15, consistent with comparative genomics.
- ADAM2 expression was found to be exclusively in the testis.
Conclusions:
- The porcine ADAM2 gene structure and size are conserved across mammalian species.
- The chromosomal localization and testis-specific expression of ADAM2 in pigs provide insights into its role in reproduction.
