PMID- 17095005
OWN - NLM
STAT- MEDLINE
DCOM- 20070213
LR  - 20061201
IS  - 0022-1759 (Print)
IS  - 0022-1759 (Linking)
VI  - 317
IP  - 1-2
DP  - 2006 Dec 20
TI  - Use of a PCR-based amplification analysis as a substitute for the Southern blot 
      method to determine the C4A and C4B genes.
PG  - 126-31
AB  - The human C4 complement components of the C4 gene are encoded by two genes, C4A 
      and C4B, located on chromosome 6p21.3 of the major histocompatibility complex 
      (MHC) of the human leukocyte antigen (HLA) class III region. Genetic 
      determination of these two genes was by the Southern blot method: the 276- and 
      191-bp NlaIV fragments represent the C4A gene with the sequence, PCPVLP, at 
      residues 1101-1106; the 467-bp NlaIV fragment represents the C4B gene with the 
      sequence, LSPVIH, at residues 1101-1106. Here, we describe a PCR-based approach 
      for differential amplification of the C4 genes adjacent to the respective 
      CYP21A1P and CYP21A2, followed by NlaIV restriction digestion in a secondary PCR 
      product and direct analysis by electrophoresis on an agarose gel to determine the 
      C4A and C4B genes. From the results of this study, we concluded that 87% and 85% 
      of the C4 genes adjacent to the CYP21A1P and CYP21A2 genes carried the C4A and 
      C4B genes, respectively. The frequencies of the C4A and C4B genes comprising the 
      C4 locus were 51.5 and 49%, respectively in this ethnic Chinese (Taiwanese) 
      cohort. Since no radiolabelling application is involved, the protocol is reliable 
      as a substitute for the Southern blot method for C4A and C4B determination.
FAU - Lee, Hsien-Hsiung
AU  - Lee HH
AD  - Department of Medical Research, Mackay Memorial Hospital, No. 45 Min-Sheng Road, 
      Tanshui, Taipei County 251, Taiwan. hhlee@ms1.mmh.org.tw
FAU - Tseng, Yung-Te
AU  - Tseng YT
FAU - Lee, Yann-Jinn
AU  - Lee YJ
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20061018
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 80295-49-4 (Complement C4a)
RN  - 80295-50-7 (Complement C4b)
SB  - IM
MH  - *Blotting, Southern/methods
MH  - Complement C4a/*genetics
MH  - Complement C4b/*genetics
MH  - Electrophoresis, Gel, Two-Dimensional
MH  - *Gene Frequency
MH  - Humans
MH  - Polymerase Chain Reaction/*methods
EDAT- 2006/11/11 09:00
MHDA- 2007/02/14 09:00
CRDT- 2006/11/11 09:00
PHST- 2006/08/11 00:00 [received]
PHST- 2006/09/13 00:00 [revised]
PHST- 2006/09/24 00:00 [accepted]
PHST- 2006/11/11 09:00 [pubmed]
PHST- 2007/02/14 09:00 [medline]
PHST- 2006/11/11 09:00 [entrez]
AID - S0022-1759(06)00272-9 [pii]
AID - 10.1016/j.jim.2006.09.018 [doi]
PST - ppublish
SO  - J Immunol Methods. 2006 Dec 20;317(1-2):126-31. doi: 10.1016/j.jim.2006.09.018. 
      Epub 2006 Oct 18.