CIS-ACTING ELEMENTS INVOLVED IN THE REGULATION OF MOUSE CLARA CELL-SPECIFIC 10-KDA PROTEIN GENE - IN-VITRO AND IN-VIVO ANALYSIS

Transient transfection and murine germ line gene transfer analysis was used to determine the regions of DNA necessary to confer the appropri ate level and cell specificity of the expression of the gene coding fo r the murine Clara cell 10-kDa protein, mCC10. To identify the cis-act ing elements involved in the regulation of mCC10 gene, different lengt hs of the B'-flanking sequence were ligated to the bacterial chloramph enicol acetyltransferase gene for transient transfection to H441 cells (human lung adenocarcinoma cell line). The corresponding sequences we re also fused to the human growth hormone gene and transferred to the murine genome for an in vivo analysis of mCC10 promoter activity, The results of the transient transfection analysis identified the region f rom -166 to -124 of the 5'-flanking region of the mCC10 gene as necess ary for the expression of this gene in H441 cells, The transgenic mous e analysis confirmed that the 166 base pairs of 5'-flanking DNA was su fficient to confer cell specific expression, However, the transgenic m ouse analysis also showed that, to achieve the full quantitative level of transgene (human growth hormone) expression, regions between -803 and -166 base pairs of the B'-flanking sequences are required for maxi mum expression of mCC10 gene promoter activity.