PMID- 8995183
OWN - NLM
STAT- MEDLINE
DCOM- 19970206
LR  - 20190830
IS  - 0093-7711 (Print)
IS  - 0093-7711 (Linking)
VI  - 45
IP  - 3
DP  - 1997
TI  - Identification of new mamu-DRB alleles using DGGE and direct sequencing.
PG  - 171-9
AB  - Rhesus macaques represent important animal models for biomedical research. The 
      ability to identify macaque major histocompatibility complex (Mhc) alleles is 
      crucial for fully understanding these models of autoimmune and infectious 
      disease. Here we describe a rapid and unambiguous way to distinguish DRB alleles 
      in the rhesus macaque using the polymerase chain reaction, denaturing gradient 
      gel electrophoresis (DGGE), and direct sequencing. The highly variable second 
      exon of Mamu-DRB alleles was amplified using generic DRB primers and alleles were 
      separated by DGGE. DNA was then reamplified from plugs removed from the gel and 
      alleles were determined using fluorescent-based sequencing. Validity of this 
      typing procedure was confirmed by identification of all DRB alleles for three 
      macaques previously characterized by cloning and sequencing techniques. 
      Importantly, our analysis revealed DRB alleles not previously identified in the 
      three reference animals. Using this technique, we identified 40 alleles in 
      fifteen unrelated macaques. On the basis of phylogenetic tree analyses, 14 new 
      DRB alleles were assigned to 10 different Mhc-DRB lineages. Interestingly, two of 
      the new DRB6 lineages had previously been identified in prosimians and pigtailed 
      macaques. Whereas traditional DRB typing methods provide limited information, our 
      new technique provides a simple and relatively rapid way of identifying DRB 
      alleles for tissue typing, determining individual identification and studies of 
      disease association and susceptibility. This new technique should also contribute 
      to ongoing studies of Mhc function and evolution in many different species of 
      nonhuman primates.
FAU - Knapp, L A
AU  - Knapp LA
AD  - Wisconsin Regional Primate Research Center, University of Wisconsin 1220 Capitol 
      Court, Madison, WI 53715, USA.
FAU - Cadavid, L F
AU  - Cadavid LF
FAU - Eberle, M E
AU  - Eberle ME
FAU - Knechtle, S J
AU  - Knechtle SJ
FAU - Bontrop, R E
AU  - Bontrop RE
FAU - Watkins, D I
AU  - Watkins DI
LA  - eng
SI  - GENBANK/U57940
SI  - GENBANK/U57941
SI  - GENBANK/U57942
SI  - GENBANK/U57943
SI  - GENBANK/U57944
SI  - GENBANK/U57945
SI  - GENBANK/U57946
SI  - GENBANK/U57947
SI  - GENBANK/U57948
SI  - GENBANK/U57949
SI  - GENBANK/U57950
SI  - GENBANK/U57951
SI  - GENBANK/U57952
SI  - GENBANK/U57953
SI  - GENBANK/U57954
SI  - GENBANK/U57955
SI  - GENBANK/U57956
SI  - GENBANK/U57957
SI  - GENBANK/U57958
SI  - GENBANK/U57959
SI  - GENBANK/U57960
GR  - AI32426/AI/NIAID NIH HHS/United States
GR  - RR00167/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Immunogenetics
JT  - Immunogenetics
JID - 0420404
RN  - 0 (HLA-DR Antigens)
SB  - IM
MH  - *Alleles
MH  - Animals
MH  - Base Sequence
MH  - Electrophoresis, Polyacrylamide Gel/methods
MH  - HLA-DR Antigens/classification/*genetics
MH  - Macaca mulatta
MH  - Molecular Sequence Data
MH  - Phylogeny
MH  - Polymerase Chain Reaction
MH  - Protein Denaturation
MH  - Sequence Analysis, DNA
EDAT- 1997/01/01 00:00
MHDA- 1997/01/01 00:01
CRDT- 1997/01/01 00:00
PHST- 1997/01/01 00:00 [pubmed]
PHST- 1997/01/01 00:01 [medline]
PHST- 1997/01/01 00:00 [entrez]
AID - 10.1007/s002510050186 [doi]
PST - ppublish
SO  - Immunogenetics. 1997;45(3):171-9. doi: 10.1007/s002510050186.