Properties of rat and mouse β-glucuronidase mRNA and cDNA, including evidence for sequence polymorphism and genetic regulation of mRNA level
| Autor: | Marc Brabant, Karen J. Moore, Jidong Cai, Gail Vander Molen, Michael R. Felder, Christopher Tietze, Gordon Watson a, Leonard Rabinow, Lynda Tussey Bracey, Cesar Labarca, Kenneth Paigen |
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| Rok vydání: | 1985 |
| Předmět: |
Mice
Inbred A Kidney law.invention Mice Sebaceous Glands Species Specificity law Complementary DNA Genetics Animals Genomic library Amino Acid Sequence RNA Messenger Cloning Molecular Gene Polymerase Glucuronidase Messenger RNA Polymorphism Genetic Base Sequence biology Hybridization probe RNA Rats Inbred Strains DNA General Medicine Molecular biology Rats Mice Inbred C57BL Genes Protein Biosynthesis Vagina biology.protein Recombinant DNA Female |
| Zdroj: | Gene. 36:15-25 |
| ISSN: | 0378-1119 |
| DOI: | 10.1016/0378-1119(85)90065-4 |
| Popis: | cDNA clones containing partial sequences for beta-glucuronidase (beta G) were constructed from rat preputial gland RNA and identified by their ability to selectively hybridize beta G mRNA. One such rat clone was used to isolate several cross-hybridizing clones from a mouse-cDNA library prepared from kidney RNA from androgen-treated animals. Together, the set of mouse clones spans about 2.0 kb of the 2.6-kb beta G mRNA. Using these cDNA clones as probes, a genomic polymorphism for DNA restriction fragment size was found that proved to be genetically linked to the beta G gene complex. A fragment of beta G cDNA was subcloned into a vector carrying an SP6 polymerase promoter to provide a template for the in vitro synthesis of single-stranded RNA complementary to beta G mRNA. This provided an extremely sensitive probe for the assay of beta G mRNA sequences. Using either nick-translated cDNA or transcribed RNA as a hybridization probe, we found that mouse beta G RNA levels are strongly induced by testosterone, and that induction by testosterone is pituitary-dependent. During the lag period preceding induction, during the induction period itself, and during deinduction following removal of testosterone, beta G mRNA levels paralleled rates of beta G synthesis previously measured by in vivo pulse-labelling experiments. Genetic variation in the extent of induction affected either the level of beta G mRNA or its efficiency of translation depending on the strain of mice tested. |
| Databáze: | OpenAIRE |
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