Held W A, Gallagher J F, Hohman C M, Kuhn N J, Sampsell B M, Hughes R G
Department of Molecular and Cellular Biology, Roswell Park Memorial Institute, Buffalo, New York 14263.
Mol Cell Biol. 1987 Oct;7(10):3705-12. doi: 10.1128/mcb.7.10.3705-3712.1987.
Mouse Ltk- cells were stably transfected with cloned genes encoding the mouse major urinary proteins (MUPs). C57BL/6J MUP genomic clones encoding MUP 2 (BL6-25 and BL6-51), MUP 3 (BL6-11 and BL6-3), and MUP 4 (BL6-42) have been identified. In C57BL/6J mice, MUP 2 and MUP 4 are known to be synthesized in male, but not female, liver, and MUP 3 is known to be synthesized in both male and female liver and mammary gland. A BALB/c genomic clone (BJ-31) was shown to encode a MUP that is slightly more basic than MUP 2 and was previously shown to be synthesized in both male and female liver of BALB/c but not C57BL/6 mice. Comigration on two-dimensional polyacrylamide gels of the MUPs encoded by the transfecting gene provides a basis for tentative identification of the tissue specificity and mode of regulation of each gene. DNA sequence analysis of the 5' flanking region indicates that the different MUP genes are highly homologous (0.20 to 2.40% divergence) within the 879 base pairs analyzed. The most prominent differences in sequence occur within an A-rich region just 5' of the TATA box. This region (from -47 to -93) contains primarily A or C(A)N nucleotides and varies from 15 to 46 nucleotides in length in the different clones.
将编码小鼠主要尿蛋白(MUPs)的克隆基因稳定转染至小鼠Ltk-细胞。已鉴定出编码MUP 2(BL6 - 25和BL6 - 51)、MUP 3(BL6 - 11和BL6 - 3)和MUP 4(BL6 - 42)的C57BL/6J MUP基因组克隆。在C57BL/6J小鼠中,已知MUP 2和MUP 4在雄性而非雌性肝脏中合成,且已知MUP 3在雄性和雌性肝脏及乳腺中均有合成。一个BALB/c基因组克隆(BJ - 31)被证明编码一种比MUP 2碱性略强的MUP,且先前已证明其在BALB/c的雄性和雌性肝脏中均有合成,但在C57BL/6小鼠中无合成。转染基因编码的MUP在二维聚丙烯酰胺凝胶上的共迁移为初步鉴定每个基因的组织特异性和调控模式提供了基础。对5'侧翼区域的DNA序列分析表明,在所分析的879个碱基对内,不同的MUP基因高度同源(差异为0.20%至2.40%)。序列中最显著的差异出现在TATA框上游仅5'处的富含A的区域。该区域(从 - 47至 - 93)主要包含A或C(A)N核苷酸,且在不同克隆中的长度从15至46个核苷酸不等。