BRIEF NOTES

Linkage mapping of the mitochondrial aconi-tase (ACO2) gene to chicken chromosome 1

T Shimogiri�, S Miyagawa†, E J Lee†, H Mannen†,S Okamoto�, Y Maeda� and S Tsuji†

*Faculty of Agriculture, Kagoshima University, Korimoto, Kago-

shima, Japan. †Faculty of Agriculture, Kobe University, Nada-ku,

Kobe, Japan.

Accepted 15 January 2002

Source/description: Mitochondrial aconitase (EC 4.2.1.3) is an

essential and ubiquitous enzyme in all animals and is linked to

the ageing process1. In mammals, the gene (ACO2) has been

mapped to HSA222, BTA5 and MMU10. However, ACO2 has

not been mapped in chickens. In order to map chicken ACO2,

we cloned cDNA using human ACO2 cDNA as a probe. The

resulting nucleotide sequence has been submitted to GenBank

(accession number AF428141).

Primer sequences: Forward: 5¢-GGCAGATGAGTTCCAGCAA-

TACTT-3¢Reverse: 5¢-TTTAAACTTCTTCCCATCTGCTCCTGTC-3¢These primers were designed to amplify the region corres-

ponding to exon 8–13 of the human ACO2 gene3.

PCR conditions: The PCR mixture contained 40 ng of genomic

DNA, 1 · PCR buffer with 2 mM MgCl2, 200 lM of each dNTP,

10 lM of each primer and 1 U of Taq DNA polymerase in a final

volume of 20 ll. PCR amplifications were performed on a

GeneAmp� PCR system 9700 (Applied Biosystems, Tokyo) with

the following profile: initial denaturation of 1 min at 94 �C,

30 cycles of 94 �C for 30 s, 65 �C for 30 s, and 72 �C for 90 s,

and a final elongation step of 7 min at 72 �C. The size of

the amplified fragment was estimated to be approximately

3580 bp. After amplification, digestion with HaeIII was carried

out for 3 h at 37 �C. Two alleles were detected on a 2% agarose

gel. Allele 1 consisted of 950, 900, 730, 420, 350 and 230 bp

fragments. Allele 2 consisted of 950, 900, 500, 420, 350 and

230 (doublet) bp fragments.

Mendelian inheritance: These two alleles demonstrated Mende-

lian inheritance in the Kobe University resource family4.

Linkage analysis and chromosomal localization: Using this marker,

segregation of the ACO2 RFLP was compared with that of about

800 genetic markers previously genotyped on this family.

Linkage analysis followed Lee et al. (2002)4. The ACO2 marker

was significantly linked to markers located on chromosome 1

(Fig. 1). Groenen et al. (2000)5 indicate a segment on chro-

mosome 1p close to where we have mapped ACO2 that con-

tains the following human chromosome 22 markers: H5,

NAGA, ADSL and MAFF. Our results suggest that ACO2 is part

of this segment (Fig. 1). Previous reports and our data suggest

that a segment of chicken chromosome 1p is syntenic with a

segment on human chromosome 22q.

Acknowledgements

We are grateful to Prof J.B. Dodgson (Michigan State Univer-

sity) for informative advice and M. Sakamoto for technical

assistance.

References1 Yan L.J. et al. (1997) Proceedings of the National Academy of

Science, USA 94, 11168–72.

2 Geurts van Kessel A.H. et al. (1980) Cytogenetics and Cell

Genetics 28, 169–72.

3 Mirel D.B. et al. (1998) Gene 213, 205–18.

4 Lee E.J. et al. (2002) Animal Genetics 33, 42–8.

5 Groenen M.A.M. et al. (2000) Genome Research 10, 137–47.

Correspondence: T Shimogiri, (simogiri@farm.agri.kagoshima-u.ac.jp)

Interspecific amplification of peccarymicrosatellite markers using porcine primers

J. Gongora�,†, Y. Chen�, J. E. Bernal†,F. W. Nicholas� and C. Moran�

*Centre for Advanced Technologies in Animal Genetics and

Reproduction, Faculty of Veterinary Science, University of Sydney,

NSW 2006, Australia. †Centro de Biologia Molecular, Gimnasio

Campestre, Calle 165 Numero 19-50, Bogota, Colombia

Accepted: 6 March 2002

Source/description: There are several examples of microsatellite

primers designed from sequences of one species being used

successfully to amplify products from other species. The groups

Figure 1 Chromosomal localization of the chicken ACO2 gene by

linkage analysis. Using PCR–RFLP, the ACO2 gene was mapped to

chicken chromosome 1 as shown by linkage analysis employing the

Kobe University resource family4. The left map contains a segment of

chromosome 1 mapped by Groenen et al. (2000)5. Loci mapped to

human chromosome 22 are boxed. Loci connecting the two maps are

underlined.

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