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本研究根据公布的锯缘青蟹SCY2基因序列设计引物SCY2-RFLPs和SCY2-RFLPa,利用PCR-RFLP技术分析了5个地理群体(宁德、汕头、万宁、东方、三亚)共计148个拟穴青蟹SCY2基因的多态性.结果表明,所扩增的SCY2基因片段含有2个StuⅠ内切酶识别位点,位于第一外显子的位点没有多态性,位于第一内含子的位点存在多态性.在148个个体中共检测到AA和AB两种基因型,等位基因B的出现频率非常低,只在三亚群体的1个个体中检测到,而在其他4个群体中均未检测到,其等位基因频率为0.003 4.序列分析结果表明,扩增片段长度为1 016 bp,包含部分5′非翻译区(282 bp)、第一外显子(131 bp)和部分第一内含子(603 bp).AA基因型表现为3条带(从大到小依次为509,356,151 bp),AB基因型表现为4条带(从大到小依次为660,509,356,151bp).B等位基因是由于867 bp处发生了C→G的单碱基突变导致StuⅠ内切酶失去了1个识别位点而形成的.
In this study, primers SCY2-RFLPs and SCY2-RFLPs were designed based on published sequences of SCY2 gene of Scylla serrata. PCR-RFLP was used to analyze 148 geographical sites (Ningde, Shantou, Wanning, Dongfang and Sanya) The results showed that the amplified SCY2 gene fragment contained two Stu Ⅰ endonuclease recognition sites, located in the first exon site without polymorphism, located in the first intron The genotypes of AA and AB were detected in 148 individuals, the frequency of allele B was very low, detected only in one individual in Sanya sub-population, while in the other four The allele frequency was 0.003 4. Sequence analysis showed that the amplified fragment was 1 016 bp in length, including a part of 5 ’untranslated region (282 bp), the first exon (131 bp ) And some of the first intron (603 bp) .AA genotype showed three bands (descending order of 509,356,151 bp), AB genotype showed four bands (descending order 660,509,356,151bp) The B allele is due to a C → G single base mutation at position 867 bp that results in the loss of one recognition site for the StuI endonuclease Into.