PCR isolation and cloning of novel splice variant mRNAs from known drug target genes.
Alternative splicing of pre-mRNAs is an important mechanism for the generation of vertebrate protein diversity. Unfortunately, the contribution of alternative splicing to protein diversity is currently not well understood because many full-length mRNA sequences have yet to be identified. In this report, we describe the use of RT-PCR to identify and clone 279 novel alternatively spliced mRNAs from 114 well-known drug target genes. Our findings demonstrate the existence of many novel alternatively spliced mRNA transcripts and suggest that many more genes undergo functionally significant alternative splicing than previously thought.
Jin
Pei
P
Incyte Corporation, 3160 Porter Drive, Palo Alto, CA 94304, USA.
Fu
Glenn K
GK
Wilson
Amy D
AD
Yang
Junming
J
Chien
David
D
Hawkins
Phillip R
PR
Au-Young
Janice
J
Stuve
Laura L
LL
D017398:Alternative Splicing; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D020224:Expressed Sequence Tags; D015894:Genome, Human; D006801:Humans; D008969:Molecular Sequence Data; D004364:Pharmaceutical Preparations; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018076:DNA, Complementary; D004364:Pharmaceutical Preparations; D012333:RNA, Messenger