Part 1 - Probe and clone availability

IGH group IGH
gene name		 Probe Clone name ATCC number HGMP number JCRB number Accession number
IGHC G3 pSgamma3h [1] 59622/59623 871 HG177 X04646
G3 pSH3Cgamma3 [1] 59624/59625 868 HG178 X03604/X03604
G3 pSH2-gamma3 [2] 59620/59621 872 HG176 X03604/X03604
Probe A pRXb1.8 [3] 61228/61229 870 HG203
Probe B pH3.9EZZ [3] 61230/61231 869 HG204
Probe C pBR0.9EZZ [3] 61236/61237 873 HG196 M25383/J03079/M29704 (1)

Part 2 - Probe and clone characteristics

Clone name Gene content Insert size (bp) Restriction sites Vector Phage or cosmid (2) Cell line (3) Comments
pSgamma3h [1] IGHG3 hinge exons (H1 to H4) 620 SacI - blunt end pUC18 lambdaEZZgamma3 [2] (4) EZZ
[4,5] (5)		 Specific G3 probe. Is the first, and so far, the only probe specific for a human Ig subclass [1]. To get the insert, digest with EcoRI and HindIII.
pSH3Cgamma3 [1] IGHG3 hinge exons (H1 to H4), CH2 and CH3 2000 SacI - SphI pUC19 lambdaEZZgamma3 [2] EZZ
[4,5] (5)		 Cross-hybridizes to all human IGHG genes. Probe used to detect RFLP of the IGHG genes [7,8].
pSH2-gamma3 [2] IGHG3 7300 HindIII pBR322 lambdaEZZgamma3 [2] EZZ
[4,5] (5)		 Contains the 7.3 kb G3 HindIII fragment from lambdaEZZgamma3 [2]. This plasmid shows some instability. We recommend growing on LB agar with ampicillin, and picking colonies to select the correct clone.
pRXb1.8 [3] Between IGHG4 and IGHE 1800 EcoRI - XbaI pUC19 cosmid cosIg23 (=cosIg10) [6] (6) Contains the EcoRI-XbaI fragment located 11.2 kb in 3' of IGHG4 and 11.8 kb in 5' of IGHE. Cross-hybridizes with highly homologous sequences present 3' to the other IGHG (G3, G1, GP and G2) genes [3].
pH3.9EZZ [3] IGHE switch sequence 3900 HindIII pUC19 lambdaepsilon10EZZ [3] EZZ
[4,5] (5)		 Contains part of the switch epsilon sequence.
pBR0.9EZZ [3] Between EZZ IGHG3 and IGHGE [3] 900 BamHI - EcoRI pUC19 lambdaEZZ4 [3] EZZ
[4,5] (5)		 This probe cross-hybridizes with highly homologous sequences present 3' to the IGHG genes except G2 (G3, G1, GP and G4) genes, which appear to be hot spots of recombination (hsg3, hsg1, hsgp and hsg4) involved in multigene deletions involving IGHG genes [3].
IMGT notes:
(1) Accession numbers corresponding to the pBR0.9cos4 clone described in [3]: M25383/J03079/M29705.
(2) Phages, or cosmids when indicated, from which have been isolated the clones used as probes.
(3) Human cell lines from which DNA was extracted for construction of the phage genomic libraries.
(4) pSgamma3h has been isolated from pSH3Cgamma3 [1] clone (2 kb insert in pUC19), itself isolated from the lambdaEZZgamma3 phage [2].
(5) EZZ and TOU [4] are TOUII-4 and TOUII-5, respectively, in the family tree [5]. EZZ (TOUII-4) EBV transformed cell line is available at ECACC (n° 93093003).
(6) cosIg10 is available at HGMP (Probe 753).
References:
[1] Huck, S. et al., FEBS Lett., 208, 221-230 (1986).
[2] Huck, S. et al., Nucl. Acids Res., 14, 1779-1789 (1986).
[3] Keyeux, G. and al., Genomics, 5, 431-441 (1989).
[4] Lefranc, M.-P. et al., Nature, 300, 760-762 (1982).
[5] Lefranc, M.-P. et al., Mol. Biol. Med., 1, 207-217 (1983).
[6] Flanagan, J. and Rabbitts, T.H., Nature, 300, 709-713 (1982).
[7] Ghanem, N. et al., Eur. J. Immunol., 18, 1059-1065 (1988).
[8] Ghanem, N. et al., Eur. J. immunol., 18, 1067-1072 (1988).