Degenerate primer design to clone the human repertoire of immunoglobulin heavy chain variable regions

ABSTRACT Immunoglobulin variable region promoters are predominantly B-cell specific, but the molecular basis for this specificity has not been elucidated. To further understand how B-cell-specific immunoglobulin promoter expression is mediated, the murine lymphoid cell line 2017 was engineered to express the green fluorescent protein under the control of an immunoglobulin heavy chain promoter and selected for high activity using multiple rounds of fluorescence-activated cell sorting. Rare clones with intense and stable immunoglobulin promoter activity were isolated. Transient transfection experiments demonstrated that two different immunoglobulin promoters and two other B-cell-specific promoters have higher activities in the selected cell lines relative to the parental line and to the non-cell-type-specific histone H2B promoter. The increased immunoglobulin activity required nucleotide residues downstream of the transcription initiation site which were also important for maximal activity in B cells and which were conserved in other B-cell-specific promoters. Unlike the unselected cells, the 2017 variants also showed activation of their endogenous immunoglobulin heavy chain variable regions.

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Effect of somatic hypermutation on potential N-glycosylation sites in human immunoglobulin heavy chain variable regions

Molecular Immunology ◽

10.1016/s0161-5890(00)00038-9 ◽

2000 ◽

Vol 37 (3-4) ◽

pp. 107-113 ◽

Cited By ~ 47

Author(s):

D Dunn-Walters

Keyword(s):

Heavy Chain ◽

Somatic Hypermutation ◽

Immunoglobulin Heavy Chain ◽

Human Immunoglobulin ◽

Variable Regions ◽

Glycosylation Sites

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Characterization of immunoglobulin heavy chain variable regions in the Mexican axolotl

Immunogenetics ◽

10.1007/bf00241261 ◽

1994 ◽

Vol 39 (3) ◽

Cited By ~ 3

Author(s):

JulienS. Fellah ◽

Caroline Jacques ◽

Jacques Charlemagne

Keyword(s):

Heavy Chain ◽

Immunoglobulin Heavy Chain ◽

Mexican Axolotl ◽

Variable Regions

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Expression of an idiotype (Id-460) during in vivo anti-dinitrophenyl antibody responses. I. Mapping of genes for Id-460 expression to the variable region of immunoglobulin heavy-chain locus and to the variable region of immunoglobulin kappa-light-chain locus.

Journal of Experimental Medicine ◽

10.1084/jem.152.3.720 ◽

1980 ◽

Vol 152 (3) ◽

pp. 720-729 ◽

Cited By ~ 22

Author(s):

E A Dzierzak ◽

C A Janeway ◽

R W Rosenstein ◽

P D Gottlieb

Keyword(s):

Heavy Chain ◽

Light Chain ◽

Regulatory Gene ◽

Variable Region ◽

Immunoglobulin Heavy Chain ◽

Kappa Light Chain ◽

Myeloma Protein ◽

Variable Regions ◽

Chain Locus

The genetic contro of the expression of an idiotype (Id-460) associated with the 2,4-dinitrophenyl (DNP)-binding BALB/c myeloma protein MOPC 460 was studied using congenic strains of mice. It was shown that the expression of high levels of Id-460 during secondary in vivo anti-DNP-ovalbumin responses was determined by genes governing immunoglobulin heavy-chain variable and kappa-light chain variable regions (V kappa). Appropriate alleles at both loci were required for the expression of Id-460. Genes in the major histocompatability complex and the X-linked immune deficiency gene found in strain CBA/N did not greatly affect Id-460 expression. The V kappa gene controlling Id-460 expression can be differentiated from Lyt-3, and it is the first instance in which expression of an idiotype subdivides the V kappa genes associated with the Lyt-3a allele. Although it is likely that the V kappa gene(s) involved are structural, the involvememt of a regulatory gene linked to the structural gene can not be excluded.

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