Use of antisense RNA to help identify a genomic clone for the 5' region of mouse beta-glucuronidase.
It was possible to gauge the inhibition of mouse beta-glucuronidase expression by injecting RNA, made from both strands of subclones of a cosmid containing the complete gene, into mouse blastomeres at the four-cell stage. Although our initial screen did not identify the 5' region, we were able to isolate a subclone containing homology to 20 bp coding for N-terminal amino acids of rat and human beta-glucuronidase structural genes. Antisense RNA prepared from one strand of the 350 bp Pst I subclone inhibited beta-glucuronidase expression by 89% while RNA prepared from the other strand had little effect. The subclone appears to correspond to the 350 bp fragment identified by others as one including the ATG start site of mouse beta-glucuronidase.
Bevilacqua
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Department of Human Genetics, University of Michigan School of Medicine, Ann Arbor 48109-0618.
Erickson
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RP
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000201:enzymology; D003001:Cloning, Molecular / Q000379:methods; D003360:Cosmids; D047108:Embryonic Development; D005260:Female; D005796:Genes; D005966:Glucuronidase / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D051379:Mice; D011247:Pregnancy; D012313:RNA; D012315:RNA Caps; D016372:RNA, Antisense; D015183:Restriction Mapping / Q000379:methods
D012315:RNA Caps; D016372:RNA, Antisense; D012313:RNA; D005966:Glucuronidase