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从100个10bp寡核苷酸引物的扩增带中,筛选出了3个RAPD特异扩增带,即团头鲂“浦江1号”群体所具有的S37277bp、S112660bp带,以及对照群体所具有的S28946bp带。该3条RAPD特异扩增带经回收、克隆和测序,并根据部分序列信息分别设计了3对各20bp的正向引物和反向引物。用3对引物分别在“浦江1号”群体和对照群体中进行PCR扩增,分别产生了Sc1(248bp)、Sc2(540bp,460bp)和Sc3(320bp)4个SCAR扩增带。其中,Sc2(540bp,460bp)和Sc3(320bp)3个SCAR扩增带在“浦江1号”和对照群体中均呈阳性,不能有效地区分这2个群体;而Sc1产生的248bp带仅在“浦江1号”群体中出现,对照群体中未出现此扩增带。采用大样本对该Sc1标记(248bp)进行验证,结果发现,在团头鲂“浦江1号”良种群体的分布频率高达91.1%(82/90)以上,而在淤泥湖原种群体中的分布频率为14.3%(8/56),Sc1标记在良种群体中的高频率,可作为检测团头鲂“浦江1号”良种的一个重要的分子遗传特征指标;另外,该Sc1标记在团头鲂“浦江1号”群体中的高出现频率,也说明了在长期(16年)和高强度(0.03%～0.04%选择率)的选育过程中,团头鲂“浦江1号”已大大提高了某些优良性状基因的等位基因频率。
Three bands of RAPD-specific amplification were screened from the amplification bands of 100bp oligonucleotide primers, that is, S37277bp and S112660bp bands contained in the “Pujiang 1” S28946bp belt. The three RAPD specific bands were recovered, cloned and sequenced. Based on the partial sequence information, three pairs of 20 bp forward and reverse primers were designed respectively. Three pairs of primers were used to amplify four SCAR bands of Sc1 (248bp), Sc2 (540bp, 460bp) and Sc3 (320bp) respectively in “Pujiang 1” population and control population. Among them, three SCAR bands of Sc2 (540bp, 460bp) and Sc3 (320bp) were positive in both “Pujiang 1” and control population, which could not effectively distinguish these two populations; however, the 248bp band generated by Sc1 was only “Pujiang No. 1” groups appear, the control population does not appear in this amplification band. The large sample was used to validate the Sc1 marker (248 bp). The results showed that the distribution frequency of elite population of “Pujiang 1” was higher than 91.1% (82/90) Frequency of 14.3% (8/56). The high frequency of Sc1 marker in elite population could be used as an important molecular genetic marker to detect elite “Pujiang 1” breed. In addition, The high frequency of occurrence in “Pujiang 1” group also shows that “Pujiang 1” has been greatly increased in the long-term (16 years) and high-intensity (0.03% ~ 0.04% selectivity) The allele frequency of some excellent trait genes.