Intragenic and intergenic sequences regulating the expression of the 5'-to-5' linked adult αand β-globin genes from large yellow croaker Pseudosciaena crocea
Department of Bioengineering and Environmental Science, Changsha University, Changsha 410003, China;Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou 310029, China
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Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou 310029, China
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Department of Bioengineering and Environmental Science, Changsha University, Changsha 410003, China
One adult α-globin gene and one β-globin gene have been cloned from the large yellow croaker Pseudosciaena crocea. Linkage analysis indicated that the αand β-globin genes were oriented head-to-head relative to each other. To identify the regulatory elements present in the intergenic and intragenic regions of the globin complex, the intergenic region alone or together with the β-globin gene first intron was cloned into the luciferase-reporter vector pGL3-Basic respectively, and the chimeric constructs were transiently transfected into Vero cells and primary fish erythrocytes. The intergenic region cannot support the high-level expression of luciferase. However, the promoter activity of the intergenic region was strongly stimulated by the positive regulatory elements (PRE) located in the β-globin gene intron 1. Thus, it is proposed that the intergenic promoters and intragenic PRE were necessary for the effective expression of the linked αand β-globin genes.