A dual-labeling method for identifying differentially expressed genes: use in the identification of cDNA clones that hybridize to RNAs whose abundance in tomato flowers is potentially regulated by gibberellins

Neil E. Olszewski; Robert T. Gast; Frederick M. Ausubel

doi:10.1016/0378-1119(89)90369-7

A dual-labeling method for identifying differentially expressed genes: use in the identification of cDNA clones that hybridize to RNAs whose abundance in tomato flowers is potentially regulated by gibberellins

Neil E. Olszewski, Robert T. Gast, Frederick M. Ausubel

Plant and Microbial Biology

Research output: Contribution to journal › Article › peer-review

16 Scopus citations

Abstract

A method for identifying cDNA clones that hybridize to differentially expressed RNAs is described. Briefly, the RNA population in which the RNAs of interest are more abundant is used as a template for the synthesis of ³⁵S-labeled cDNAs and another RNA population in which the RNAs of interest are less abundant is used as a template for the synthesis of ³²P-labeled cDNAs. The labeled cDNAs are pooled and hybridized to plaque or colony lifts constructed from a cDNA library. Clones that hybridize to RNAs that are differentially expressed are identified using differential autoradiography/fluorography to discriminate between the ³²P and ³⁵S isotopes. We have used this method to identify cDNA clones that hybridize to mRNAs that are more abundant in the flowers of wild-type tomato than in the flowers of mutants that have low endogenous levels of gibberellins.

Original language	English (US)
Pages (from-to)	155-162
Number of pages	8
Journal	Gene
Volume	77
Issue number	1
DOIs	https://doi.org/10.1016/0378-1119(89)90369-7
State	Published - Apr 15 1989

Bibliographical note

Funding Information:
We thank J. Berman for making suggestionst hat improved the manuscript. This work was supported by research funds from the Graduate School of the University of Minnesota and a grant from Hoechst AG to Massachusetts General Hospital, R.T.G. is a

Keywords

Lycopersicon esculentum
Recombinant DNA
colony hybridization
differential autoradiography/fluorography
flower development
gib-1 mutant
molecular cloning
plaque hybridization

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A dual-labeling method for identifying differentially expressed genes: use in the identification of cDNA clones that hybridize to RNAs whose abundance in tomato flowers is potentially regulated by gibberellins. / Olszewski, Neil E.; Gast, Robert T.; Ausubel, Frederick M.
In: Gene, Vol. 77, No. 1, 15.04.1989, p. 155-162.

Research output: Contribution to journal › Article › peer-review

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title = "A dual-labeling method for identifying differentially expressed genes: use in the identification of cDNA clones that hybridize to RNAs whose abundance in tomato flowers is potentially regulated by gibberellins",

abstract = "A method for identifying cDNA clones that hybridize to differentially expressed RNAs is described. Briefly, the RNA population in which the RNAs of interest are more abundant is used as a template for the synthesis of 35S-labeled cDNAs and another RNA population in which the RNAs of interest are less abundant is used as a template for the synthesis of 32P-labeled cDNAs. The labeled cDNAs are pooled and hybridized to plaque or colony lifts constructed from a cDNA library. Clones that hybridize to RNAs that are differentially expressed are identified using differential autoradiography/fluorography to discriminate between the 32P and 35S isotopes. We have used this method to identify cDNA clones that hybridize to mRNAs that are more abundant in the flowers of wild-type tomato than in the flowers of mutants that have low endogenous levels of gibberellins.",

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author = "Olszewski, {Neil E.} and Gast, {Robert T.} and Ausubel, {Frederick M.}",

note = "Funding Information: We thank J. Berman for making suggestionst hat improved the manuscript. This work was supported by research funds from the Graduate School of the University of Minnesota and a grant from Hoechst AG to Massachusetts General Hospital, R.T.G. is a",

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A dual-labeling method for identifying differentially expressed genes: use in the identification of cDNA clones that hybridize to RNAs whose abundance in tomato flowers is potentially regulated by gibberellins

Abstract

Bibliographical note

Keywords

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