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初级精母细胞中的Cre表达:一种用于生殖系基因工程的工具。

Cre expression in primary spermatocytes: a tool for genetic engineering of the germ line.

作者信息

Vidal F, Sage J, Cuzin F, Rassoulzadegan M

机构信息

Unité 470 de l'Institut National de la Santé et de la Recherche Médicale, Université de Nice, France.

出版信息

Mol Reprod Dev. 1998 Nov;51(3):274-80. doi: 10.1002/(SICI)1098-2795(199811)51:3<274::AID-MRD6>3.0.CO;2-M.

Abstract

Transgenic mice were generated expressing a testicular Cre recombinase driven by promoter sequences derived from the gene encoding Synaptonemal Complex Protein 1 (Sycp1), expressed at an early stage of the male meiosis (leptotene to zygotene). Recombination at target LoxP sites was examined during germinal differentiation in mice harboring Sycp1-Cre and a second transgene where LoxP sites flank either the beta geo coding region, the Pgk1 promoter, or a tk-neo cassette inserted into the Rxr alpha locus. The LoxP-flanked transgenes were stably maintained in the somatic tissues of the double transgenic animals, as well as in the progeny of the females. Mice born after mating the double-transgenic males with normal females showed extensive deletions of the LoxP-flanked sequences. When the males were hemizygous for the Sycp1-Cre transgene, the deletions were observed even in the fraction of the offspring which had not inherited the Cre gene, thus demonstrating that expression occurred in the male parent during spermatogenesis. The high efficiency of excision at the LoxP sites makes the Sycp1-Cre transgenic males suitable for evaluating the role of defined gene functions in the germinal differentiation process.

摘要

构建了转基因小鼠,其表达由编码联会复合体蛋白1(Sycp1)的基因启动子序列驱动的睾丸Cre重组酶,该蛋白在雄性减数分裂早期(细线期到偶线期)表达。在携带Sycp1-Cre和第二个转基因的小鼠生殖细胞分化过程中,检测了靶LoxP位点的重组情况,第二个转基因中LoxP位点位于β-geo编码区、Pgk1启动子或插入RXRα基因座的tk-neo盒两侧。LoxP侧翼的转基因在双转基因动物的体细胞组织以及雌性后代中稳定维持。将双转基因雄性与正常雌性交配后出生的小鼠显示出LoxP侧翼序列的广泛缺失。当雄性对Sycp1-Cre转基因为半合子时,即使在未继承Cre基因的部分后代中也观察到缺失,从而证明在精子发生过程中雄性亲本中发生了表达。LoxP位点的高效切除使得Sycp1-Cre转基因雄性适合用于评估特定基因功能在生殖细胞分化过程中的作用。

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