Wirgin I I, Maceda L, Mesing C
Institute of Environmental Medicine, New York University Medical Center, Tuxedo 10987.
J Hered. 1992 Sep-Oct;83(5):375-82. doi: 10.1093/oxfordjournals.jhered.a111234.
We tested the ability of cellular oncogene (c-onc) probes to identify F1 hybrids and the lineage of known backcrosses within the fish genus Morone. Total DNA was isolated from five to 14 individuals per North American Morone species (striped bass, white bass, white perch, and yellow bass). The DNA was digested with two restriction enzymes, Eco RI and Hin dIII, Southern blotted, and hybridized to six different c-onc probes including v-abl, v-erb B, c-myc, c-H-ras, c-K-ras, and v-src. We found fixed genotypic differences among the four species for all six probes in single restriction enzyme digests. The heritability of these nuclear DNA genotypes was evaluated in hatchery-produced F1 Morone hybrids (striped bass x white bass and striped bass x white perch) tested with the six informative single probe/restriction enzyme combinations. All F1 individuals exhibited heterozygosity in all diagnostic nuclear DNA fragments, confirming the Mendelian inheritance of these genotypes in these fish. Furthermore, analysis of these nuclear DNA genotypes in hatchery-produced backcrosses of F1 hybrids striped bass x (white bass x striped bass) detected both recombinant and parental genotypes at all six polymorphic c-onc sequences. The lineage of suspected Morone hybrids of unknown descent collected from Lewis Smith Lake, Alabama, and from the Occoquan River, Virginia, was determined using the c-onc probes. Our results suggest that c-onc probes are suitable markers to unequivocally identify F1 hybrids and backcrosses and to quantify introgression in natural populations of fishes. The addition of RFLP analysis of mtDNA provided a complete ancestral history of individual fish.
我们测试了细胞癌基因(c-onc)探针识别鱼类马苏大麻哈鱼属内F1杂种以及已知回交谱系的能力。从北美马苏大麻哈鱼属的每个物种(条纹鲈、白鲈、白鲈和黄鲈)的5至14个个体中分离出总DNA。用两种限制性内切酶Eco RI和Hin dIII消化DNA,进行Southern印迹,然后与六种不同的c-onc探针杂交,包括v-abl、v-erb B、c-myc、c-H-ras、c-K-ras和v-src。我们发现,在单限制性内切酶消化中,所有六种探针在这四个物种之间存在固定的基因型差异。用六种信息丰富的单探针/限制性内切酶组合对孵化场生产的F1马苏大麻哈鱼杂种(条纹鲈×白鲈和条纹鲈×白鲈)进行测试,评估了这些核DNA基因型的遗传力。所有F1个体在所有诊断性核DNA片段中均表现出杂合性,证实了这些基因型在这些鱼类中的孟德尔遗传。此外,对F1杂种条纹鲈×(白鲈×条纹鲈)孵化场生产的回交后代的这些核DNA基因型进行分析,在所有六个多态性c-onc序列中均检测到重组基因型和亲本基因型。使用c-onc探针确定了从阿拉巴马州刘易斯·史密斯湖和弗吉尼亚州奥科坎河采集的血统不明的疑似马苏大麻哈鱼杂种的谱系。我们的结果表明,c-onc探针是明确识别F1杂种和回交以及量化鱼类自然种群基因渗入的合适标记。线粒体DNA的RFLP分析补充提供了单个鱼类完整的祖先历史。