PMID- 10648862
OWN - NLM
STAT- MEDLINE
DCOM- 20000501
LR  - 20190723
IS  - 0022-1759 (Print)
IS  - 0022-1759 (Linking)
VI  - 233
IP  - 1-2
DP  - 2000 Jan 13
TI  - Rapid cloning of HLA class I cDNAs by locus specific PCR.
PG  - 119-29
AB  - The human major histocompatibility complex (MHC) class I loci, human leukocyte 
      antigen (HLA)-A, -B, -C, encode highly polymorphic molecules that mediate immune 
      recognition of infectious pathogens and can initiate the rapid rejection of 
      transplanted tissue. Cloning of HLA class I alleles is complicated by 
      polymorphism as well as interlocus homology. Here, HLA class I cDNAs are 
      amplified by PCR using one common primer with one of three locus specific primers 
      whose 3' ends map to conserved, locus specific nucleotides. Using these primers, 
      HLA-A, -B, and -C alleles were cloned from a number of cell lines and two 
      different HLA-B alleles were cloned from a single, heterozygous cell line. The 
      amplified products encode the entire extracellular portion of the class I 
      molecules. An amplified HLA-A allele was cloned into an expression vector and the 
      protein product was detected on the surface of a transfected cell. A premature 
      termination codon was engineered into the HLA-A allele by site directed 
      mutagenesis and the soluble protein product was detected in the culture medium of 
      transfected cells. Therefore, these primers can be used to rapidly clone, alter, 
      and express HLA class I molecules. This method may expedite the generation of 
      reagents for testing the antigen specificity of antibodies, natural killer cells, 
      or T cells.
FAU - Johnson, D R
AU  - Johnson DR
AD  - Department of Pathology, 454 Boyer Center for Molecular Medicine, Yale University 
      School of Medicine, 295 Congress Avenue, New Haven, CT, USA. 
      david.johnson@yale.edu
FAU - Biedermann, B C
AU  - Biedermann BC
FAU - Mook-Kanamori, B
AU  - Mook-Kanamori B
LA  - eng
GR  - R01-HL43364/HL/NHLBI NIH HHS/United States
GR  - R29-AI35099/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Complementary)
RN  - 0 (HLA Antigens)
RN  - 0 (HLA-A Antigens)
RN  - 0 (HLA-B Antigens)
RN  - 0 (HLA-C Antigens)
SB  - IM
MH  - Alleles
MH  - Base Sequence
MH  - Cell Membrane/immunology
MH  - Cells, Cultured
MH  - Cloning, Molecular/*methods
MH  - DNA Primers/genetics
MH  - DNA, Complementary/*genetics
MH  - Evaluation Studies as Topic
MH  - Genes, MHC Class I
MH  - HLA Antigens/*genetics
MH  - HLA-A Antigens/genetics
MH  - HLA-B Antigens/genetics
MH  - HLA-C Antigens/genetics
MH  - Humans
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction/*methods
MH  - Solubility
MH  - Transfection
EDAT- 2000/01/29 09:00
MHDA- 2000/05/08 09:00
CRDT- 2000/01/29 09:00
PHST- 2000/01/29 09:00 [pubmed]
PHST- 2000/05/08 09:00 [medline]
PHST- 2000/01/29 09:00 [entrez]
AID - S0022175999001210 [pii]
AID - 10.1016/s0022-1759(99)00121-0 [doi]
PST - ppublish
SO  - J Immunol Methods. 2000 Jan 13;233(1-2):119-29. doi: 
      10.1016/s0022-1759(99)00121-0.