Transcription of cloned tRNA gene fragments and subfragments injected into the oocyte nucleus of Xenopus laevis.
Cloned 3.18 kilobase fragments of Xenopus laevis DNA containing genes for tRNAMet1 and for at least one other 4S RNA species are transcribed rapidly after their injection into the nucleus of X. laevis oocytes. The newly synthesized RNA can be resolved by gel electrophoresis into a few predominant 4S RNA species and a series of slower migrating components. One of the 4S RNA species appears to be identical, by fingerprint analysis, to the tRNAMet1 isolated by hybridization of somatic cell RNA to this cloned tRNA gene fragment (tDNA). Thus, the tRNAMet1 produced after injection can be both fully processed and modified. Its rate of synthesis is estimated to be about 6 x 10(9) molecules/hr in each oocyte injected with 2 ng of tDNA. When the tDNA fragment is cleaved into two halves with restriction endonuclease Sst I, each injected half gives rise to a subset of the RNAs produced after injection of the intact fragment. This experiment thus suggests the presence of at least two transcriptional units on this cloned tDNA. This simple way of biologically testing defined restriction fragments may be of value for analyzing the functional organization of other cloned eukaryotic DNA units.
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D003090:Coliphages / Q000235:genetics; D004247:DNA / Q000008:administration & dosage; D004274:DNA, Recombinant; D005260:Female; D005796:Genes; D008715:Methionine; D008845:Microinjections; D008970:Molecular Weight; D009865:Oocytes / Q000378:metabolism; D012343:RNA, Transfer / Q000235:genetics*; D014158:Transcription, Genetic; D014981:Xenopus / Q000235:genetics
D004274:DNA, Recombinant; D004247:DNA; D012343:RNA, Transfer; D008715:Methionine
Proc Natl Acad Sci U S A. 1977 Apr;74(4):1502-6
193103
Proc Natl Acad Sci U S A. 1977 May;74(5):2064-8
266727
Proc Natl Acad Sci U S A. 1977 Feb;74(2):560-4
265521
Cell. 1977 Jul;11(3):663-71
560257
Cell. 1976 Jun;8(2):183-95
986875
J Biol Chem. 1973 Jul 25;248(14):5024-32
4577761
J Morphol. 1972 Feb;136(2):153-79
4109871
Annu Rev Biochem. 1975;44:273-93
166604
Eur J Biochem. 1975 Sep 1;57(1):85-96
1175643
Proc Natl Acad Sci U S A. 1974 Jul;71(7):2900-4
4527857
Dev Biol. 1973 Sep;34(1):106-18
4787598
Biochem J. 1972 Apr;127(3):597-605
4672799
Biochemistry. 1968 Feb;7(2):668-74
4868544
Proc Natl Acad Sci U S A. 1971 Dec;68(12):3175-9
5289255
J Mol Biol. 1975 May 25;94(3):441-8
1100841
Eur J Biochem. 1975 Dec 1;60(1):295-302
1204643
J Embryol Exp Morphol. 1968 Nov;20(3):401-14
5750105
Natl Cancer Inst Monogr. 1966 Dec;23:431-47
5963987
Nucleic Acids Res. 1974 Mar;1(3):331-53
10793670