Molecular cloning and sequence analysis of heavy- and light-chain variable region genes of anti-CD71 monoclonal antibody.
OBJECTIVE
To clone heavy-chain and light-chain variable region (VH and VL) gene of mouse-anti-human CD71 monoclonal antibody (mAb).


METHODS
One-step method was used to extract total RNA, and a set of oligonucleotide primers were designed to amplify the cDNAs with reverse transcriptase-polymerase chain reaction (RT-PCR), and the resultant products were respectively cloned into PMD18-T vector and their sequences analyzed.


RESULTS
The PCR product obtained with the oligonucleotide primers for the variable region of mouse immunoglobulin heavy chain was about 350 bp and that with oligonucleotide primers for the light chain was about 320 bp, and their DNA sequences were determined.


CONCLUSIONS
The length of the cloned heavy chain variable region was 348 bp, belonging to mouse heavy-chain subgroup II(A); the light-chain variable region was 336 bp that belongs to mouse kappa light-chain subgroup II.
Wang
Zhi-You
ZY
Department of Immunology, Institute of Biological Products of Wuhan, Wuhan 430060, China.
Shen
Tao
T
Zhang
Ai-Hua
AH
Bi
Lan
L
Jin
Yu-Ling
YL
Sun
Ke-Fang
KF
Yu
Mo-Song
MS
Shi
Liang-Ru
LR
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D011990:Receptors, Transferrin; D017422:Sequence Analysis, DNA
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C068126:CD71 antigen; D018076:DNA, Complementary; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D011990:Receptors, Transferrin