Abstract: Objective To investigate the optimal breeding method and genotype identification method of RELM-β gene knockout rats. Method The RELM-β - / - homozygous female Sprague Dawley ( SD) rats and wild-type male Wild type (WT) SD rats were mated according to 2 ∶ 1 to produce F1 heterozygous rats. Four mating method, RELM-β - / - homozygous cross, RELM-β + / - heterozygous cross, homozygous and heterozygous cross ( orthogonal and reverse cross) , were used to identify the genotype of offspring rats by PCR amplification gel electrophoresis. The appearance of the offspring rats was observed, and the number, weight and homozygous rate of the offspring rats were calculated. Result The genotypes of the parents and offspring rats were successfully identified by PCR amplification gel electrophoresis. There was no significant difference in the appearance and morphology of the three genotypes of rats, no significant difference in the body weight of the rats aged from 3 weeks to 6 weeks, and no significant difference in the number of offspring of different mating method. Conclusion Correct reproduction and identification is an important way to obtain RELM-β gene knockout rats. PCR amplification gel electrophoresis is rapid, economical and reproducible in identifying rat genotypes.

Key words: gene knockout, resistin like molecule-β, genotypic identification, breeding methods

摘要： 目的 探讨抵抗素样分子 β(RELM-β)基因敲除大鼠的最优繁育方式和基因型鉴定方法。 方法 将 RELMβ- / -纯合子雌性 Sprague Dawley( SD)大鼠和野生型雄性 Wild type( WT) SD 大鼠按 2 ∶ 1 交配,繁育出 F1 杂合子大鼠,采用 RELM-β- / -纯合子互交、RELM-β+ / -杂合子互交、纯合子及杂合子互交( 正交及反交) 4 种方式交配,PCR 扩增凝胶电泳法鉴定子代大鼠基因型,观察子代大鼠的外形,统计子代大鼠数量、体质量及纯合率。 结果 PCR 扩增凝胶电泳法成功鉴定亲代及子代大鼠的基因型,3 种基因型大鼠的外观形态无明显差异,3 周至 6 周龄大鼠体质量无明显差异,不同交配方式子代数量无明显差异。 结论 合理的繁殖方法并进行正确的鉴定是获得 RELM-β 基因敲除 SD 大鼠的重要途径,PCR 扩增凝胶电泳法具有快速、经济,重复性较好的优点。

关键词: 基因敲除, 抵抗素样分子 β, 基因型鉴定, 繁育方法