Isolation and characterization of human blood platelet mRNA and construction of a cDNA library in lambda gt11. Confirmation of the platelet derivation by identification of GPIb coding mRNA and cloning of a GPIb coding cDNA insert.
We have developed a purification method for the isolation of platelet-specific poly (A+) RNA and demonstrated that human blood platelets, despite the absence of a nucleus, contain stable mRNA. The poly (A+) RNA was used to construct a platelet-specific cDNA expression library in lambda gt11. The platelet derivation of the purified mRNA was confirmed by identification of membrane glycoprotein Ib (GPIb) message by immunoprecipitation of rabbit reticulocyte lysate translation products with poly- and monoclonal antibodies against GPIb alpha and by sequencing of a GPIb alpha cDNA clone.
Wicki
A N
AN
Theodor Kocher Institute, Bern, Switzerland.
Walz
A
A
Gerber-Huber
S N
SN
Wenger
R H
RH
Vornhagen
R
R
Clemetson
K J
KJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D001792:Blood Platelets / Q000032:analysis* / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000302:isolation & purification*; D005796:Genes; D015698:Genomic Library; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D010980:Platelet Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D012333:RNA, Messenger / Q000097:blood / Q000302:isolation & purification*
D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger; D004247:DNA