[Cloning and sequence analysis of the light chain variable region gene of monoclonal anti-idiotypic antibody NP30 of Schistosoma japonicum].
OBJECTIVE
To amplify and sequence the light chain of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum.


METHODS
By comparing the conserved regions at each end of the nucleotide sequences of murine germ-line genes encoding FR1 and FR4 regions of immunoglobulin light chain variable regions, we designed a set of primers for amplification of VL gene. The hybridoma cells secreting anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum were cultured and their genome DNAs were extracted and used as templates for PCR. The PCR product was then cloned into pUC19 vector. The recombinants were sequenced by Sanger's method. The VL gene was compared with GenBank and published mouse VL genes.


RESULTS
The full-length of VL gene was 318 bp. The VL gene was a member of mouse Ig kappa light chain subgroup IV and generated from rearrangement of germ line V and J kappa 4 genes. The VL gene sequence has been registered by GenBank(accession No. AF206720).


CONCLUSIONS
The obtained VL gene was a potentially functional gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum.
Song
X T
XT
Institute for Molecular Biology of Medicine, Nanjing Medical University, Nanjing 210029.
Feng
Z Q
ZQ
Qiu
Z N
ZN
Li
Y Q
YQ
Yu
X C
XC
Xiong
Y
Y
Yin
C C
CC
Huang
H L
HL
Guan
X H
XH
D000595:Amino Acid Sequence; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000235:genetics*; D000911:Antibodies, Monoclonal / Q000235:genetics*; D003001:Cloning, Molecular; D005784:Gene Amplification; D005803:Genes, Immunoglobulin; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008969:Molecular Sequence Data; D012549:Schistosoma japonicum / Q000276:immunology*; D017422:Sequence Analysis, DNA
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region