An improved shotgun antisense method for mutagenesis and gene identification. Academic Article

abstract

• Shotgun expression of antisense cDNA, where each transformed cell expresses a different antisense cDNA, has been used for mutagenesis and gene identification in Dictyostelium discoideum. However, the method has two limitations. First, there were too few clones in the shotgun antisense cDNA library to have an antisense cDNA for every gene in the genome. Second, the unequal transcription level of genes resulted in many antisense cDNAs in the library for some genes but relatively few antisense cDNAs for other genes. Here we report an improved method for generating a larger antisense cDNA library with a reduced percentage of cDNA clones from highly prevalent mRNAs and demonstrate its utility by screening for signal transduction pathway components in D. discoideum.

authors

• Gomer, Richard

published proceedings

• Biotechniques

altmetric score

• 3.2

author list (cited authors)

• Tang, Y. u., & Gomer, R. H.

citation count

• 1

complete list of authors

• Tang, Yu||Gomer, Richard H

publication date

• January 2020

publisher

• Future Science Group  Publisher

published in

• BioTechniques: the journal of laboratory technology for bioresearch  Journal

keywords

• Cdna Normalization
• DNA, Antisense
• DNA, Complementary
• DNA, Protozoan
• Dictyostelium
• Genetic Screen
• Mutagenesis
• Sequence Analysis, DNA
• Shotgun Antisense

PubMed Central ID

• 31973564

Digital Object Identifier (DOI)

• 10.2144/btn-2019-0123

start page

• 163

end page

• 165

volume

• 68

issue

• 3

URL

• http://dx.doi.org/10.2144/btn-2019-0123