1982Proceedings of the National Academy of SciencesOpen access

Heavy chain genes of rabbit IgG: isolation of a cDNA encoding gamma heavy chain and identification of two genomic C gamma genes.

Christine Martens, Kevin W. Moore, Michael Steinmetz, Leroy Hood, K L Knight

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Abstract

A cDNA library was constructed by using rabbit spleen poly(A)+RNA as template, and from this library was isolated a cDNA clone, p2a2, that encodes 179 amino acids of the heavy chain of rabbit IgG. The nucleotide sequence of p2a2 showed that it encodes the COOH-terminal eight amino acids of the CH1 domain, the hinge region, the CH2 domain, and the NH2-terminal half of the CH3 domain of C gamma. Southern blot hybridization analysis of rabbit sperm DNA showed that two EcoRI fragments hybridized strongly with the C gamma cDNA. The p2a2 cDNA was used as a probe to isolate recombinant Charon 4A phage clones containing C gamma sequences from a genomic library of rabbit liver DNA. Two distinct DNA segments were identified by restriction mapping and hybridization analysis, suggesting that the haploid rabbit genome may contain two different C gamma genes.

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A cDNA library was constructed by using rabbit spleen poly(A)+RNA as template, and from this library was isolated a cDNA clone, p2a2, that encodes 179 amino acids of the heavy chain of rabbit IgG. The nucleotide sequence of p2a2 showed that it encodes the COOH-terminal eight amino acids of the CH1 domain, the hinge region, the CH2 domain, and the NH2-terminal half of the CH3 domain of C gamma. Southern blot hybridization analysis of rabbit sperm DNA showed that two EcoRI fragments hybridized strongly with the C gamma cDNA. The p2a2 cDNA was used as a probe to isolate recombinant Charon 4A phage clones containing C gamma sequences from a genomic library of rabbit liver DNA. Two distinct DNA segments were identified by restriction mapping and hybridization analysis, suggesting that the haploid rabbit genome may contain two different C gamma genes.

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Available abstract

A cDNA library was constructed by using rabbit spleen poly(A)+RNA as template, and from this library was isolated a cDNA clone, p2a2, that encodes 179 amino acids of the heavy chain of rabbit IgG. The nucleotide sequence of p2a2 showed that it encodes the COOH-terminal eight amino acids of the CH1 domain, the hinge region, the CH2 domain, and the NH2-terminal half of the CH3 domain of C gamma. Southern blot hybridization analysis of rabbit sperm DNA showed that two EcoRI fragments hybridized strongly with the C gamma cDNA. The p2a2 cDNA was used as a probe to isolate recombinant Charon 4A phage clones containing C gamma sequences from a genomic library of rabbit liver DNA. Two distinct DNA segments were identified by restriction mapping and hybridization analysis, suggesting that the haploid rabbit genome may contain two different C gamma genes.

Key concepts: Complementary DNA, Molecular biology, Southern blot, Biology, cDNA library, genomic DNA, EcoRI, Genomic library

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Heavy chain genes of rabbit IgG: isolation of a cDNA encoding gamma heavy chain and identification of two genomic C gamma genes. — Research Paper | ScholarLens