Cloning and analysis of the untranslated regions of the Xenopus laevis Connexin30 mRNA.
The full-length 5' and 3' untranslated regions (UTRs) of Xenopus laevis Connexin30 (Cx30) mRNA were cloned and sequenced. The Cx30 messenger contains a 148 nt 5' UTR and a 480 nt 3' UTR. Four different constructs were made to enable the analysis of the role of the Cx30 UTRs in translation efficiency and in protein localization in the early Xenopus embryo. Transcripts encoded the Green Fluorescent Protein (GFP) reporter and contained the 5' and 3' UTR of either Cx30 or globin. In vivo analyses after injection of the transcripts into one cell-stage Xenopus embryos showed that the Cx30 3' UTR enables very efficient translation. The 5' UTR was slightly inhibitory compared with the globin 5' UTR. The localization of the produced GFP was analyzed. GFP was ubiquitously expressed in all parts of the embryo. Based on this observation we conclude that neither the 5' UTR nor the 3' UTR confers specific localization of the translation of the Cx30 mRNA in the embryo.
Meijer
H A
HA
Department of Developmental Biology, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.
Dictus
W J
WJ
Thomas
A A
AA
D020121:5' Untranslated Regions / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D000075302:Connexin 30; D017630:Connexins / Q000235:genetics*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D029867:Xenopus Proteins; D014982:Xenopus laevis / Q000196:embryology / Q000235:genetics*; D000097002:Gap Junction beta-1 Protein
D020121:5' Untranslated Regions; D000075302:Connexin 30; D017630:Connexins; D018076:DNA, Complementary; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D029867:Xenopus Proteins; D000097002:Gap Junction beta-1 Protein