NDLI: The effect of different 3′ untranslated regions on the accumulation and stability of transcripts of a gfp transgene in chloroplasts of transplastomic tobacco

Content Provider	Springer Nature Link
Author	Tangphatsornruang, Sithichoke Birch Machin, Ian Newell, Christine A. Gray, John C.
Copyright Year	2010
Abstract	The 3′ untranslated region (3′ UTR) of transcripts is a major determinant of transcript stability in plastids and plays an important role in regulating gene expression. In order to compare the effect of different 3′ UTRs on transgene expression in tobacco chloroplasts, the 3′ UTRs from the tobacco chloroplast rbcL, psbA, petD and rpoA genes and the terminator region of the Escherichia coli rrnB operon were inserted downstream of the gfp reporter gene under the control of the psbA promoter, and the constructs were introduced into the plastid genome by particle bombardment. RNA-gel blot analysis of homoplasmic transplastomic plants identified gfp transcripts of ~1.0 and ~1.4 kb from all constructs and showed that plants expressing gfp with the rrnB terminator contained 4 times more gfp transcripts than plants expressing gfp with the rbcL and rpoA 3′ UTRs. The amounts of transcripts accumulated roughly correlated with the half-life of the transcripts, determined by RNA-gel blot analysis of transcripts present in leaves treated with actinomycin D to prevent continued transcription of the chimeric gfp genes. Transcripts containing the 3′ region of rrnB were most stable, with half-lives of ~43 h, considerably longer than the half-lives of the other ~1.0 kb gfp transcripts (13–26 h). Immunoblot analysis with antibodies to GFP indicated that all plants contained about the same amount of GFP (~0.2% total soluble protein), suggesting either that translation was limited by something other than the amount of transcript or that the 3′ UTR was affecting translation.
Starting Page	385
Ending Page	396
Page Count	12
File Format	PDF
ISSN	01674412
Journal	Plant Molecular Biology
Volume Number	76
Issue Number	3-5
e-ISSN	15735028
Language	English
Publisher	Springer Netherlands
Publisher Date	2010-09-22
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Chloroplast GFP RNA stability Tobacco Transplastomic plant 3′ Untranslated region Biochemistry Plant Sciences Plant Pathology
Content Type	Text
Resource Type	Article
Subject	Genetics Plant Science Medicine Agronomy and Crop Science

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