Rapid identification and isolation of zebrafish cDNA clones

Rapid identification and isolation of zebrafish cDNA clones

A fast and economical approach, referred to as cDNA clone tagging, was adapted to identify and isolate zebrafish cDNA clones. The basic approach was to partially sequence the coding region of size selected cDNA clones and the partial sequences were then used as tags for identifying the clones throug...

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Bibliographic Details
Published in:Gene Vol. 201; no. 1; pp. 87 - 98
Main Authors: Gong, Zhiyuan, Yan, Tie, Liao, Ji, Lee, Sze Ern, He, Jiangyan, Hew, Choy L
Format: Journal Article
Language:English
Published: Netherlands Elsevier B.V 12-11-1997
Subjects:
Animals
Base Sequence
Danio rerio
DNA, Complementary
Embryo
EST
Gene
Genome
Molecular Sequence Data
Teleost
Zebrafish - genetics
Teleost
Embryo
Gene
YAC, yeast artificial chromosome
EST
kb, kilobase(s)
PCR, polymerase chain reaction
BAC, bacterial artificial chromosome
bp, base pair(s)
Genome
DANA, Danio retroposon A
PAC, P1 artificial chromosome
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Description
Summary:A fast and economical approach, referred to as cDNA clone tagging, was adapted to identify and isolate zebrafish cDNA clones. The basic approach was to partially sequence the coding region of size selected cDNA clones and the partial sequences were then used as tags for identifying the clones through homology search. To benefit maximally from the tagging approach, two cDNA libraries, derived from embryonic and adult fish poly(A) + RNAs, respectively, were constructed by unidirectional cloning; conceptually, they have the potential to represent all expressed zebrafish genes. A total of 1084 clones were sequenced from the two libraries, and 511 clones were identified, based on sequence homology. These identified clones were derived from at least 261 genes, encoding 48 translational machinery proteins, 47 cytosolic proteins, 43 cytoskeletal proteins, 41 nuclear proteins, 32 membrane proteins, 22 secreted proteins, 20 mitochondrial proteins and 8 proteins with an unknown location. Of the 261 distinct cDNA clones identified, 254 were isolated for the first time in the zebrafish. These tagged cDNA clones, identified and unidentified, provide rich resources for developmental analysis as well as mapping of zebrafish genome. The long-term objective of this study is to establish a tagged zebrafish gene library that can be accessed both by hybridization screening against the plasmid DNAs and by electronic screening using the sequence information.
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content type line 23
ISSN:0378-1119
1879-0038
DOI:10.1016/S0378-1119(97)00431-9