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GrainGenes Reference Report: PMB-23-643

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Reference	PMB-23-643
Title	Transcient and stable expression of gusA fusions with rice genes in rice, barley and perennial ryegrass
Journal	Plant Molecular Biology
Year	1993
Volume	23
Pages	643-669
Author	Hensgens L [ Show all 10 ]
Abstract	Transcriptional and translational fusions were made between the reading frame coding for beta-D-glucuronidase and sequences of either a constitutively expressed rice gene (GOS2) involved in initiation of translation or a light-inducible rice gene (GOS5) The transient expression of the fusions was studied via particle bombardment of seedling tissues of rice, perennial ryegrass and barley Furthermore, the results of transient and stable expression were compared for cell suspensions of four rice varieties, one barley variety and one perennial ryegrass variety. The GOS2-gusA fusions were active in all three monocots studied. Best results were obtained for a construct having both a transcriptional and a translational fusion as well as intron and exon sequences (PORCEHyg). The level of GUS activity was in the range of activities as obtained by the 35S CaMV promoter transcriptionally fused to gusA. The gusA fusion with the light-inducible gene (GOS5) was active in green seedling tissues of all monocots studied. Also a weak expression compared to the GOS2 constructs was found in stably transformed rice callus. The gusA fusions with the mannopine synthase promoters 1' and 2' of the T(R)-DNA were transiently expressed at lower levels in cell suspensions than PORCEHyg. For stably transformed rice callus the expression of the GOS2-gusA fusion often decreased during prolonged subculture. This decrease in GUS activity and the various GUS-staining phenotypes of transgenic calli are explained by the presence of. different cell types in the suspensions used and in the calli. It is presumed that the nature of the cells and their relative contribution in the calli change drastically upon further subculture.
Keyword	[ Hide all but 1 of 18 ] beta-glucuronidase biolistic-transformation callus gene expression genetic transformation gus2 gene gus5 gene gusa gene histoenzymology hordeum vulgare introns lolium perenne oryza sativa plant proteins promoters recombinant-dna reporter genes transgenic-plants

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