外源基因在蓝太阳鱼胚胎发育过程中的表达

Foreign Gene Expression in Green Sunfish, Lepomis cyanellus During Embryonic Development

通过基因重组,将石斑鱼生长激素基因编码序列克隆到鲤鱼β＿actin基因启动子下游,构建成全鱼基因重组子。采用显微注射法将报告基因GFP基因导入蓝太阳鱼受精卵中,研究了外源基因在鱼类胚胎发育中的整合与表达的时序。结果表明GFP在原肠末期开始表达,一直到鱼苗期并未出现表达量的衰减,并表现为嵌合性表达。在受体胚胎早期,外源基因的表达与基因构型和注射的剂量有关,环形质粒的表达要略高于线形质粒,较高浓度的外源基因的表达效率要高。同时将GFP基因与全鱼基因重组子进行了共转移,结果表明两种独立的基因在受体中的整合或表达没有必然的联系。

The coding sequence of grouper Epinephelus coioides growth hormone gene was cloned under the regulation of Carp β-actin gene promoter and enhancer, so a all-fish recombinant pCAecGHc was constructed. GFP gene was microinjected into eggs of green sunfish Lepomis cyanellus in order to investigate the integration and expression of a foreign gene in host fish during embryonic development. The results indicated that GFP gene expression began at the end of gastrula stage, and lasted to fry at a stable expression level. GFP gene expression was relative to the configuration and dosage of foreign gene. In addition, GFP gene expression of mosaic fashion was observed. At the same time, co-injection of GFP gene with a growth hormone gene construct into it was conducted, showing that the integration or expression of one foreign gene into host genome cannot suggests the integration or expression of the other gene.