Linkage analysis: The HaeIII RFLP was mapped by linkage

analysis in a subset of a Large White – Meishan population5,

including the Roslin contribution to the PiGMaP pedigrees6.

The GLUL HaeIII RFLP was linked to SW174 (H ¼ 0.15;

LOD ¼ 3.84) and in a multipoint analysis to the interval be-

tween S0295 and SW174 at the distal end of SSC9.

Radiation hybrid (RH) mapping: The RH mapping was accom-

plished using the INRA-University of Minnesota porcine RH

panel (IMpRH)7,8. The PCR with Pair 2 primers was used to

screen a panel of 90 hybrid clones, and analysis of the data was

performed as described earlier9. The porcine GLUL gene was

mapped to chromosome 9. The most significant linkage

was with SW174 (40 cR; LOD ¼ 7.69) and SW2093 (90 cR;

LOD ¼ 2.66).

FISH assignment: A recombinant plasmid containing the

�2 kb insert was labelled with biotin-14-dATP by nick

translation (Gibco BRL, Uxbridge, UK) and used for standard

fluorescence in situ hybridization (FISH)10. Immunodetection

and amplification was performed using avidin-FITC and anti-

avidin biotin. Chromosomes were counterstained with propi-

dium iodide and DAPI (Sigma, St Louis, MO, USA). The G-like

pattern generated by DAPI staining was used for chromosome

identification and for regional assignment. Twenty-one (28%)

of the 74 metaphases analysed showed symmetrical double

spots on at least one copy of chromosome 9q24-q25 (Fig. 2).

No paired signal was repeatedly detected on any other chro-

mosomal region.

Comments: GLUL is located on the human sequence map on

chromosome 1 at 185.5 Mbp, in band 1q24.3 (http://

www.ensembl.org; 26/02/2002). The porcine homologue of

PTGS2 (human sequence map location ¼ 190.2 Mbp; band

1q25.2) is already known to map to SSC911. However, the

porcine homologues of ATP1B1 and SERPINC1 (AT3) (human

sequence map locations ¼ 171.6 Mbp, band 1q23.2 and

176.3 Mbp, band 1q23.3, respectively) map to SSC4. Thus the

assignment of GLUL to porcine chromosome 9q24-q25 indi-

cates that the evolutionary breakpoint is situated between

cytogenetic bands q23.2 and q24.3 of human chromosome 1,

between ATP1B1 (which is on SSC4q13-q2112) and GLUL.

Acknowledgements: We greatly appreciate Drs Martine Yerle

and Denis Milan (INRA, Castanet-Tolosan, France) for provi-

ding the IMpRH panel, and Professor Hermann Geldermann

(University Hohenheim, Stuttgart, Germany) for providing the

DNA samples from the Hohenheim pedigree material. We thank

Marie Datlova for technical assistance. This work was suppor-

ted by Grant Agency of the Czech Republic (Grant no. 523/00/

0669) and Grant Agency of the Ministry of Agriculture of the

Czech Republic (MZE-M03-99-1).

References1 Wang Y. et al. (1996) Genomics 37, 195–9.

2 Helou K. et al. (1997) Mamm Genome 8, 362–4.

3 van de Zande L. et al. (1990) Gene 87, 225–32.

4 Geldermann H. et al. (1996) J Anim Breed Genet 113, 381–7.

5 Walling G.A. et al. (1998) Anim Genet 29, 415–24.

6 Archibald A.L. et al. (1995) Mamm Genome 6, 157–75.

7 Yerle M. et al. (1998) Cytogenet Cell Genet 82, 182–8.

8 Hawken R.J. et al. (1999) Mamm Genome 10, 824–30.

9 Stratil A. et al. (2001) Anim Genet 32, 110–2.

10 Trask B.J. (1991) Method Cell Biol 35, 3–35.

11 Gladney C.D. et al. (1999) J Anim Sci 77, 787–8.

12 Lahbib-Mansais Y. et al. (1993) Genomics 15, 91–7.

Correspondence: A. Stratil (stratil@iapg.cas.cz)

Linkage relationships for 35 new microsatelliteloci in chinook salmon Oncorhynchustshawytscha

K. A. Naish and L. K. Park

Northwest Fisheries Science Center, National Marine Fisheries

Service, NOAA, Seattle, WA, USA

Accepted 26 April 2002

Source/description: Chinook salmon genomic DNA was restric-

ted, size selected for the 200–900 bp range by gel electro-

phoresis and ligated into the EcoRV site of a plasmid vector,

PZeRo 2.2 (Invitrogen, Carlsbad, CA, USA). Clones containing

microsatellite sequences were identified by screening the library

using the following [33P] end-labelled probes: [dCA]10, [dGA]10,

[dAAT]10, [dAAAT]6, [dAAG]8, [dAAC]8, [dATC]8, [dACT]8,

[dGATA]6 and [dGACA]6. Plamids from positive colonies were

isolated and the inserts were sequenced and analysed on an

ABI-377 sequencer (Thetagen, Seattle, WA, USA). Primer pairs

for each locus were developed for the flanking sequences using

Generunner V 3.0 (Hastings Software).

Figure 2 Fluorescence in situ hybridization with the porcine GLUL

plasmid clone to a porcine metaphase. Double signals on both

chromosomes 9q24-q25 are shown.

� 2002 International Society for Animal Genetics, Animal Genetics, 33, 312–327

316 Brief notes

Tab

le1

Des

crip

tion

of

prim

erse

quen

ces

use

dto

amplif

ym

icro

sate

llite

loci

inch

inook

salm

on,

Onco

rhyn

chus

tshaw

ytsc

ha.

Nom

encl

ature

of

each

of

the

loci

follo

ws

the

conve

ntion

outlin

edin

Jack

son

etal

.3The

locu

sid

entifier

ispre

ceded

by

the

spec

ies

des

ignat

ion

(Ots¼

O.

tshaw

ytsc

ha)

and

follo

wed

by

the

sourc

ela

bora

tory

(NW

FSC¼

Nort

hw

est

Fish

erie

sSc

ience

Cen

ter)

.

Locu

snam

eG

enBan

kac

cess

ion

no.

Forw

ard

prim

erse

quen

ce5¢–

3¢

Rev

erse

prim

erse

quen

ce5¢–

3¢

Tm

Size

of

cloned

alle

leSi

zera

nge

Rep

eat

clas

s

Ots

500

NW

FSC

AY

042690

AA

CTC

CTG

GA

CA

AA

CC

TC

GTG

AC

CC

TG

CC

CA

TA

AC

AC

54

240

180–2

60

GA

Ots

501

NW

FSC

AY

042691

TTTC

ATC

AC

ATC

AG

CA

GC

TG

TA

CTC

GG

TTTC

ATTTG

ATC

54

144

140–1

70

GA

Ots

502

NW

FSC

AY

042692

GTA

ATG

TG

GG

AA

GA

GTTG

GTTA

TC

CC

TTTC

TC

TTTC

TC

TC

54

132

130–1

40

GA

Ots

503

NW

FSC

AY

042693

ATC

CC

TG

TTC

TC

CTC

TTTA

CC

ATTG

CA

CA

CC

AC

AC

ATA

C54

154

150–1

70

GA

Ots

504

NW

FSC

AY

042694

GA

AA

GA

GC

GA

GA

GG

GA

GTG

AA

CA

GG

GA

AA

TG

CC

AA

ATC

54

140

140–1

60

GA

Ots

505

NW

FSC

AY

042695

GTC

TA

GC

AA

TG

ATC

AA

CA

AC

CTG

GG

AC

CA

AC

AG

TTTA

CG

54

222

220–2

30

GA

Ots

506

NW

FSC

AY

042696

GA

AG

GA

TA

GA

CG

ATG

TG

TG

CTG

CA

AC

ATA

ATA

GA

CA

CTC

CC

54

102

90–1

20

GA

Ots

507

NW

FSC

AY

042697

AG

AG

AG

GA

CTA

GG

AA

TG

GG

TC

AC

TC

TC

AC

TA

AC

TC

TG

CC

54

213

210–2

20

GA

Ots

508

NW

FSC

AY

042698

AG

GTG

TC

TG

CC

CTG

AG

AG

CA

TA

GTTG

AG

CC

ATTG

GG

54

270

320–3

50

GA

Ots

509

NW

FSC

AY

042699

GG

GA

TG

GG

AG

TTTA

CC

TC

GTTC

TTC

AA

ATTC

AA

AC

TG

G57

356

350–3

70

GTT/A

TT

Ots

510

NW

FSC

AY

042700

AC

TG

GG

AG

CTTA

TTG

TTC

AC

AC

GA

TA

AG

AG

GC

AA

AG

GA

C54

149

120–1

60

GA

Ots

511

NW

FSC

AY

042701

GG

TTTG

TG

ATTA

GC

GTG

AA

GC

CTC

AG

CC

AG

AA

CA

GA

GC

54

151

130–1

90

GA

Ots

512

NW

FSC

AY

042702

TTC

CA

TG

CC

AA

TA

AA

GC

CC

TA

CC

AC

AC

CC

TG

TC

AG

AA

G54

180

180–2

50

GA

Ots

513

NW

FSC

AY

042703

CC

CC

AG

CC

CA

AC

TA

GA

AC

GG

CC

AA

GG

AG

CTA

GG

GG

A54

297

270–3

10

GA

Ots

514

NW

FSC

AY

042704

CTG

TC

TTTC

CTC

TTTG

ATG

GA

AC

CA

GC

CA

CTA

CA

CA

C54

140

140–1

70

CA

Ots

515

NW

FSC

AY

042705

AC

AG

TG

ATG

GA

GC

TTG

ATT

CA

CG

ATTTC

TA

TTTG

TC

TC

CG

54

198

190–2

40

CA

Ots

516

NW

FSC

AY

042706

TA

CG

GTTA

TC

TC

TC

AA

AG

CTTG

AC

ATA

TTG

TG

TG

TG

ATG

54

310

200–3

20

GA

/CA

Ots

517

NW

FSC

AY

042707

TA

TTC

AC

AG

CA

CA

AC

AG

GA

CC

TG

CG

CA

CTG

ATTTA

TTA

TA

G54

212

170–2

30

GA

Ots

518

NW

FSC

AY

042708

CTA

TC

TC

CC

TC

GC

AA

CTA

AC

GC

CC

TTTG

AA

TTG

TA

GA

GA

G54

195

190–1

10

GA

Ots

519

NW

FSC

AY

042709

ATTG

AG

AG

AG

AA

AG

ATA

GA

AG

CG

TA

CC

AC

AG

AC

ATA

AA

AC

AC

TG

54

198

90–1

05

GA

Ots

520

NW

FSC

AY

042710

AG

AG

TG

CA

AG

GC

GA

GTA

TTC

CTTG

AC

AG

CA

GG

TA

AC

CA

TG

54

205

190–2

30

GA

Ots

521

NW

FSC

AY

042711

AA

GA

GA

AA

GTG

TG

AG

GG

AG

AG

AG

GG

AA

ATTA

GG

AG

AC

CA

AG

54

171

150–1

70

GA

Ots

522

NW

FSC

AY

042712

GA

AA

GG

AA

ATG

GA

GG

AA

AC

CC

CA

GA

GTTC

AG

TG

TTA

TC

TC

58

116

110–1

40

GA

Ots

523

NW

FSC

AY

042713

GTA

CG

AG

AG

AG

AG

AA

CA

TG

AG

GTA

AC

GG

AG

GG

AA

TG

ATG

C54

128

110–2

20

GA

Ots

524

NW

FSC

AY

042714

GG

CC

TA

CA

AA

CTTG

AG

TC

AG

GTG

AG

AC

AG

GA

AG

TG

GC

AG

54

142

140–1

90

GA

/TA

/GA

Ots

525

NW

FSC

AY

042715

TTTC

AA

CC

TTG

CC

CTC

TG

ATTC

AG

TG

ATA

CC

AG

GA

GA

GT

C54

128

130–1

90

GA

Ots

526

NW

FSC

AY

042716

CA

GC

AG

GA

AG

ATG

ATG

AA

GA

CA

AC

TC

CTA

ATG

GC

AG

ATA

C54

178

170–1

90

GA

Ots

527

NW

FSC

AY

042717

ATG

AG

AC

CG

CC

TG

TA

AA

CA

AC

AC

ATC

AA

TA

AC

AC

TG

TC

TG

54

144

140–1

80

GA

Ots

528

NW

FSC

AY

042718

CC

ATG

GC

AG

TTTC

GA

TA

GTG

GTC

AG

TTG

AC

TA

ATC

GC

54

111

115–1

90

GA

TA

/GA

Ots

529

NW

FSC

AY

042719

CTG

GTC

AA

AC

GC

TC

ATC

CC

CA

AC

AC

CA

CTTTC

CA

TC

AG

54

170

120–1

90

GA

Ots

530

NW

FSC

AY

042720

GG

GTTA

GTC

AC

AG

AG

GTC

AG

GG

TTA

GG

GTTG

ATA

GA

AG

GA

C54

106

100–1

40

GA

Ots

531

NW

FSC

AY

042721

TA

CG

CC

AG

GA

GA

AA

GA

CG

TC

CA

CTC

TG

TA

GC

CTTG

AC

C54

136

120–1

40

GA

Ots

532

NW

FSC

AY

042722

TC

ATTA

TC

TG

ATTTA

CTA

CA

CA

GTTA

TG

CC

TG

GTC

TG

GA

AC

54

171

90–1

20

CA

/GA

/GA

AA

Ots

533

NW

FSC

AY

042723

TTC

AC

CTC

CTTC

CA

TC

TTTC

TTA

ATG

GG

TG

TC

TG

AC

TA

TG

G54

120

100–1

20

GA

Ots

534

NW

FSC

AY

042724

TC

TG

TG

ATTTC

CC

TG

TG

CA

GA

CC

ATG

GA

TG

AC

ATC

ATC

54

104

100–1

10

GA

� 2002 International Society for Animal Genetics, Animal Genetics, 33, 312–327

317Brief notes

PCR conditions: Loci were amplified in 5 ll volumes with the

following components: 20–100 ng DNA, 10 mM Tris–HCl

(pH 9.0), 50 mM KCl, 2.0 mM MgCl2, 0.02 mM each dNTP

(Promega, Madison, WI, USA), 0.2 pmol of primer and 0.5 U of

Taq polymerase (Promega). Amplification was performed in

384-well plates using an MJ Research thermocycler; one cycle

of denaturation at 95 �C for 5 min, followed by 30–35 cycles of

95 �C for 45 s, the annealing temperature (Table 1) for 45 s

and extension at 72 �C for 1 min. PCR products were electro-

phoresed through 8% acrylamide denaturing gels on Owl Sci-

entific Penguin units (plate size 20 · 20 cm) and visualized by

silver staining (Promega).

Polymorphisms: DNA from 12 chinook salmon were used for

polymorphism screening. Six parents were derived from adult

spawners returning to the Puget Sound, Washington, USA, and

six were derived from returnees to Alaska, USA. Allele sizes

were estimated by comparison to a 10-bp ladder (Invitrogen).

Linkage data: Two outbred (heterozygous) crosses were created

for mapping studies. The choice of parents for each cross was

based on prior information on family-specific growth rates in a

quantitative genetic experiment1. Unrelated males from fast-

growing families were mated with unrelated females from

slow-growing families (one male with three females and one

male with two). All offspring were raised in a common envi-

ronment, and their growth rates were characterized at several

points over a year following the methodology of Hard et al.1

DNA material from both crosses is available on application.

Pairwise linkage relationships between marker loci were

examined in the F1 of the smaller of the two crosses, using

141 offspring from the first female and 51 offspring from

the second. Recombination frequencies were calculated using

LINKMFE5 (R. G. Danzmann, http://www.uoguelph.ca/~rdan-

zman/). Recombination rates differ between female and male

salmonids2, and recombination frequencies are reported sep-

arately for each sex (Table 2).

References1 Hard J.J. et al. (1999) J Hered 90, 597–606.

2 Sakamoto T. et al. (2000) Genetics 155, 1331–45.

3 Jackson T.R. et al. (1998) Heredity 80, 143–51.

Correspondence: K A Naish (knaish@u.washington.edu)

Current address: K. A. Naish, School of Aquatic and Fishery Sciences,

University of Washington, 1122 NE Boat St, Seattle, WA 98105, USA.

Fine mapping1 of the bovine solute carrierfamily 25, member 4 (SLC25A4) gene toBTA27q14-q15 by fluorescence in situhybridization and radiation hybrid mapping

C. Drogemuller*, H. Kuiper*, G. Hauke*,J. L. Williams† and O. Distl*

Table 2 Two-point linkage relationships with published salmonid microsatellite and allozyme loci (see Sakamoto et al.2 for details on the loci).

Omy ¼ O. mykiss, Str ¼ Salmo trutta.

Female/male marker Chinook loci Published loci Recombination frequency LOD

Female Ots 502 NWFSC OmyRGT08 TUF 0.307 4.642

Female Ots 506 NWFSC Ots522 NWFSC 0.224 3.417

Female Ots 522 NWFSC OmyRGT09 TUF 0.03 15.975

Female Ots 524 NWFSC Str073 INRA 0.208 10.508

Female Ots 529 NWFSC OmyRGT14 TUF 0.333 3.32

Female Ots 529 NWFSC OmyRGT31-02 TUF 0.083 23.029

Male Ots 500 NWFSC OmyRGT20 TUF 0.193 5.434

Male Ots 500 NWFSC OmyRGT39 TUF 0.189 12.33

Male Ots 500 NWFSC Str060 INRA 0.18 12.769

Male Ots 501 NWFSC Ots507 NWFSC 0.338 3.221

Male Ots 504 NWFSC Ots531 NWFSC 0.257 7.03

Male Ots 507 NWFSC OmyRGT21 TUF 0.33 3.519

Male Ots 508 NWFSC SSOD-1 0.139 5.397

Male Ots 509 NWFSC OmyRGT12 TUF 0 15.051

Male Ots 518 NWFSC Ots 520 NWFSC 0.074 25.157

Male Ots 518 NWFSC Str02 INRA 0.07 24.387

Male Ots 520 NWFSC Str02 INRA 0.145 15.02

Male Ots 521 NWFSC Ots534 NWFSC 0.229 3.228

Male Ots 524 NWFSC Ots534 NWFSC 0.122 6.838

Male Ots 529 NWFSC OmyRGT31-02 TUF 0.03 31.663

1 This is a more precise localization of SLC25A4 previously mapped to

BTA27 by Li and Womack (1997) Mamm Genome 8, 773–4.

� 2002 International Society for Animal Genetics, Animal Genetics, 33, 312–327

318 Brief notes