Abstract: 【Objective】The objective of the study is to confirm the two fragments of introns 7 and 8 of rat RVLG (rat vasa-like gene) pre-mRNA might be alternatively spliced in the ovary, and to further reveal the potential alternative splicing isoforms of introns 7 and 8 of RVLG pre-mRNA both in ovary and testis. In this study the existence of alternative splicing in RVLG pre-mRNA will be further confirmed and the potential alternative splicing isoforms of introns 7 and 8 of RVLG pre-mRNA both in ovary and testis will be tested. 【Method】 The cDNA template was prepared by reverse transcription of total RNA extracted from the ovaries of 5 female rats and the testes of 5 male rats. Partial coding region of 620-690 bp including the initiation codon were amplified by PCR. Fourteen independently amplified fragments, 7 each from ovary and testis, were sequenced and aligned with the genomic DNA of RVLG gene to analyze the alternative splicing pattern. 【Result】 In all the 14 sequenced cDNA, two fragments, one 45 bp and another 15 bp, were found to be involved in alternative splicing patterns. Of the 7 from ovary, 5 retained the 45 bp but lost the 15 bp, 2 retained the 15 bp but lost the 45 bp. Four from testes retained the 45 bp but lost the 15 bp, 2 retained both the 45 bp and the 15 bp, and one without the 45 bp and the 15 bp. The 45 bp was identified to locate at the 5’ end of intron 7 flanked by two GT, and the 15 bp at the 3’ end of the intron 8 flanked with two AG, granting these two fragments cryptic splice site for their alternative splicing.【Conclusion】There are multiple combinations of alternative splicing patterns of introns 7 and 8 of RVLG. The in vivo canonical splicing isoform may be the one that exon 8 with the 45 bp from intron 7, but exon 9 without the 15 bp from intron 8.

Key words: RVLG , RT-PCR , alternative splicing