Amos, Boston U, USA Kitzis, CHU-Paris, France

Barker, U Alabama Birm, USA Klinger, Integ Genet, USA

Barranger, Los Angeles, USA Knight, London, England

Barton, Cambridge, England Lavinha, Lisboa Codex, Portugal

Beaudet, Baylor, USA Lissens, Vrije U Brussels, Belgium

Boué, Paris, France Loukopoulos, Athens, Greece

Bowcock, Stanford, USA Lucotte, College de France

Cao, U Cagliari, Italy Malcolm, ICH-London, England

Carbonara, Torino, Italy Malik, Basler-Basel, Switzerland

Cassiman, U Leuven, Belgium Mao, Collab Res, USA

Claustres, Montpellier, France McIntosh, WGH-Edinburgh, Scotland

Collins, U Michigan, USA Morel, Lyon, France

Cutting, Johns Hopkins, USA Morgan, McGill, Canada

Dallapiccola, Roma, Italy Naylor, UT San Antonio, USA

De Arce, Dublin, Ireland Olek, U Bonn, West Germany

Dean, NCI Frederick, USA Orr, U Minnesota, USA

Desnick, Mount Sinai, New York, USA Pignatti, U Verona, Italy

Edwards, Oxford, England Ramsay, SAMIR, South Africa

Elles, St Mary's-Manchester, England Richards, GeneScreen, USA

Erlich, Cetus, USA Romeo, Gaslini-Genoa, Italy

Estivill, Barcelona, Spain Rowley, Rochester, USA

Ferec, Brest, France Rozen, Montreal Children, Canada

Ferrari, Milano, Italy Scheffer,UGroningen,TheNetherlands

Godet, Villeurbanna, France Schmidtke, IHG, Berlin

Goossens, Creteil, France Schwartz, U Copenhagen, Denmark

Graham, Belfast, N Ireland Sebastio, Naples, Italy

Gruenert, UCSF, USA Seltzer, U Colorado, USA

Halley, Rotterdam, The Netherlands Spona, Vienna, Austria

Harris, Guy's-London, England Super, Royal Manchester, England

Highsmith, NC Mem Hosp, USA Thibodeau, Rochester, USA

Horst, Münster, West Germany Tümmler, Hannova, West Germany

Jaume-Roig, Son Dureta, Spain Verellen-Dumoulin,Bruxelles,Belgium

Kalaydjieva, Sofia, Bulgaria Willems, U Antwerp, Belgium

Kant, U Penn, USA Williamson,St Mary'sLondon,England





NEWSLETTER #16, May 4, 1990


1. M. Dean, M.B. White, B. Gerrard and C. Stewart report the sequence of 2 more mutations (1154insTC and 1214delT) in exon 7; both are frameshift mutations found in single families. Since the quality of the original fax is poor (I told Mike but he has not sent another one), the 2 mutations are briefly summarized below. I have also taken the liberty of changing the name of the 2 to conform with the proposed nomenclature scheme. These mutations were detected by SSCP gel electrophoresis of the PCR products generated by using the previously reported primers CF34 and CF35. The 1154insTC mutation is an insertion of 2 bp (TC) after nucleotide position 1154 and it may be detected by heteroduplexes on polyacrylamide gel following digestion with Msp I or Dde I; 1214delT is a deletion of 1 bp (T) at 1214 and it destroys an Nla III site. The 2 mutations were not found on 225 other non-[[Delta]]F CF chromosomes. There is no haplotype information. Mike is willing to provide PCR primers and ASOs; the original letter is attached.

2. Kerem, Zielenski, Bozon and Tsui report the identification of 3 additional mutations:

(a) 3659delC- a frameshift mutation in exon 19 in a single CF chromosome (Toronto family #2); deletion of C at nucleotide position 3659 or 3960; haplotype IIa; not present in 57 non-[[Delta]]F508 CF chromosomes (7 from IIa) and 50 N chromosomes (43 from IIa); the deletion may be detected by PCR with a common oligonucleotide primer 19i-5 (see below) and 2 ASO primers, HSC8 (5'-GTA TGG TTT GGT TGA CTT GG-3') for the normal and HSC9 (5'-GTA TGG TTT GGT TGA CTT GT-3') for the mutant allele; the PCR condition is as usual except the annealing temperature is at 60[[ring]]C to improve specificity;

(b) 556delA- a frameshift mutation in exon 4 in a single CF chromosome (Toronto family #17, GM1076); deletion of A at nucleotide position 556; haplotype IIIb; not found in 31 other CF chromosomes (9 from IIIb) and 30 N chromosomes (16 from IIIb); the mutation creates a Bgl I site; PCR primers are 4i-5 and 4i-3 (see below); the enzyme cuts the mutant PCR product (437 bp) into 2 fragments of 287 and 150 bp in size;

(c) 621+1G->T- a single bp change affecting the splice site (GT->TT) at the 3' end of exon 4; this mutation is detected in 5 French-Canadian CF chromosomes (one each in Toronto families #22, 23, 26, 36 and 53) but not in 33 other CF chromosomes (18 from the same group, group I) and 29 N chromosomes (13 from group I); the mutation creates a Mse I site; genomic DNA may be amplified by the 2 intron primers, 4i-5 and 4i-3, and cut with MseI to distinquish the N and mutant alleles; the normal would give 4 fragments of 33, 35, 71 and 298 bp in size; the 298 bp fragment in the mutant is cleaved by the enzyme to give a 54 and a 244 bp fragments.

3. The exon/intron boundary sequences for exon 4, 7, 19 and 22, provided by Julian Zielenski (Tsui), are attached. The revised sequences for exons 10, 11, 14a, 15, 20 and 21 are also attached (we apologize for the errors). The rest will come soon.

4. The new FAX number for Cassiman is (32) 16-21.59.97.

Best regards,