Further characterization of HLA homozygous typing cell lines at the LMP2 polymorphic codon 60 by an ARMS typing method.
LMP2 is a subunit of the 20S proteasome within the cellular cytosolic compartment that is thought to cleave proteins into approximately 9 amino acid long oligopeptides. It is hypothesized that changes in the low molecular mass protease (LMP) gene sequence may alter the activity or specificity in which the LMP genes cleave peptides. Currently, the typing method for LMP2 involves polymerase chain reaction (PCR), restriction enzyme digestion, and gel electrophoresis. To help reduce the cost and cumbersomeness of this method, a new typing method was adapted for the LMP2 gene. To establish this new amplification refractory mutation system (ARMS) typing method, primers have been defined, amplification conditions optimized, and control cell lines sequenced to validate testing parameters. Results are listed for selected 10th and 11th International Histocompatibility Workshop homozygous cell lines.
Hopkins
L M
LM
Immunohematology and Serology Laboratory, Michigan State University, East Lansing 48824, USA.
Bull
P J
PJ
Gerlach
J A
JA
Bull
R W
RW
D002460:Cell Line; D003062:Codon / Q000235:genetics*; D003546:Cysteine Endopeptidases; D017931:DNA Primers; D005838:Genotype; D006680:HLA Antigens / Q000235:genetics*; D006650:Histocompatibility Testing / Q000379:methods*; D006720:Homozygote; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D011506:Proteins / Q000235:genetics*
D003062:Codon; D017931:DNA Primers; D006680:HLA Antigens; D011506:Proteins; C071313:LMP-2 protein; D003546:Cysteine Endopeptidases