[Cloning of human B lymphocyte activation-related novel gene].
OBJECTIVE
To clone the novel activation-related gene of B lymphocyte.


METHODS
The differential display reversal transcription PCR (DDRT-PCR) technique was applied to analyse the expression difference of mRNA between resting and activated B lymphocyte from human tonsil. The positive differential display cDNA fragment identified by Northern-blotting was chosen as probe to filtrate human activated B lymphocyte cDNA library.


RESULTS
Sixty two differential display cDNA fragments (expressed sequence tag, EST) were obtained. Thirty-two of them were mainly expressed in resting B lymphocyte and thirty were expressed in activated cells. Twenty-five were positive ones after identification by Northern blot analysis. A novel cDNA clone was obtained after using EST30 as a probe to filtrate the human activated B cell cDNA library. The whole cDNA clone was 2,048 bp in length and contains a 630 bp open reading frame. The N end of the deduced amino acid sequence was homologous with KAR3 protein which is a member of kinesins superfamily in yeast.


CONCLUSIONS
A novel possible activation-related gene in human B lymphocyte was obtained.
Lu
X W
XW
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China. luxingwu@263.net
Yin
J Y
JY
Cui
L X
LX
D000595:Amino Acid Sequence; D001402:B-Lymphocytes / Q000276:immunology*; D001483:Base Sequence; D002478:Cells, Cultured; D002648:Child; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D008213:Lymphocyte Activation / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D012333:RNA, Messenger