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cDNA clones encoding immunoglobulin lambda chains from rabbit expressing the phenotype c7
D J Hayzer1, G O Young-Cooper, R G Mage
1Department of Medical Biochemistry, Medical Center of the University of Geneva, Switzerland.
Insights
Rabbit immunoglobulin lambda light chains with c7 and c21 markers represent distinct isotypic forms. Gene C lambda 6 encodes the constant region of c7 lambda chains, differing from the c21-encoding gene C lambda 5.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- Rabbit immunoglobulin lambda light chains exhibit distinct phenotypes, including c7 and c21.
- Understanding the genetic basis of these isotypic forms is crucial for B cell receptor diversity.
Purpose of the Study:
- To investigate the genetic basis of rabbit immunoglobulin lambda light chain c7 and c21 phenotypes.
- To identify the specific genes encoding the constant regions of these lambda chains.
Main Methods:
- Screening of a rabbit spleen cDNA library using V lambda and C lambda probes.
- Nucleotide sequencing of hybridizing clones to determine V lambda, J lambda, and C lambda regions.
- Comparative analysis of gene sequences and Southern blotting.
Main Results:
- Three hybridizing clones showed identical V lambda, J lambda, and C lambda regions.
- The V lambda region demonstrated 97% similarity to the germ-line gene V lambda 2.
- The C lambda region was identical to the newly identified gene C lambda 6, which differs from C lambda 5 (encoding c21).
Conclusions:
- Gene C lambda 6 encodes the constant region of c7 lambda chains.
- The c7 and c21 markers designate two distinct isotypic forms of rabbit lambda light chains.
- A model is proposed to explain the genetic basis of c7 and c21 phenotypes based on Southern blotting and restriction mapping.
Abstract:
A cDNA library derived from spleen cells of an unimmunized rabbit expressing the c7 phenotype of Ig lambda chains (c7+, c21-) was screened with V lambda or C lambda probes of a lambda light chain bearing c21 epitopes. The nucleotide sequences of three hybridizing clones were found to be identical within the V lambda, J lambda and C lambda regions. The V lambda region was 97% similar to that of the functional germ-line gene V lambda 2, and the C lambda region was identical to that of gene C lambda 6, recently identified. Gene C lambda 6 exhibited four codon differences when compared with gene C lambda 5, the latter encoding c21 epitopes. The data presented here and in the accompanying report (Jaton, J.-C. et al., Eur. J. Immunol. 1990, 20:2713) support the view that gene C lambda 6 encodes the C region of c7 lambda chains and that c7 and c21 markers designate two distinct isotypic forms of lambda chains. On the basis of comparative Southern blotting analyses and restriction maps of cloned genomic regions containing V lambda and C lambda genes, a scheme is proposed to account for the c7- and c21- phenotypes.
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