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199406, Санкт-Петербург, ул.Гаванская, д. 49, корп.2

ISSN 1999-6314

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ТОМ 4, СТ. X (сc. X) // Апрель, 2003 г.

ALLELE FREQUENCY DISTRIBUTIONS OF YNZ22 AND IG-JH IN THE LITHUANIAN POPULATION

G. Chvatovic
Human Genetics Centre of Vilnius University Hospital's,
Santariskiu Clinics



Abstract

  << Contents

 

Allele and genotype frequencies for the YNZ22 and Ig-JH loci have been determined in a Lithuanian population samples using the polymerase chain reaction (PCR) amplification. In 215 unrelated individuals of YNZ22 locus, 12 different alleles and 49 different genotypes were observed. For Ig-JH locus in 200 individuals samples a total of 24 different genotypes and 11 alleles were observed.

Introduction

In human genome contain a large number of DNA segments of tandemly repeated sequences as VNTR (Variable number of tandem repeats) loci.

The technique of amplification "in vitro" using the polymerase chain reaction (PCR) has been applied successfully to amplify several VNTR. The presence of large number of alleles at VNTR loci also makes them useful in the context of paternity testing, forensic medicine and population genetics.

In order to obtain population data in the Lithuania, allele frequencies and genotype distribution of two polymorphic loci (YNZ22 and Ig-JH). The Ig- JH VNTR locus, situated on chromosome 14, the polymorphism detected using PCR consists of 13 alleles, ranging in length from 420 bp to 1020 bp with a 50 bp core sequence. The VNTR locus detected previously by probe YNZ22 on 17p13.3 chromosome is considered a useful marker. The polymorphism detected using PCR consists of 14 alleles, ranging in length from 170 bp to 1080 bp with a 70 bp core sequence.

Material and methods

DNA samples were collected from blood samples in the Lithuania corresponding to parents of families involved in paternity analysis. DNA was extracted from fresh blood leukocytes using Chelex extraction method.

DNA samples from 215 individuals were used for analysing the YNZ22 polymorphism. For the Ig- JH study 200 individuals DNA samples were typed.

The amplification was carried out with the following primer sequences and conditions:

1. YNZ22

5' AAA CTG CAG AGA GAA AGG TCG AAG AGT GAA GTG

5' AAA GGA TCC CCC ACA TCC GCT CCC CAA GTT

2. Ig-JH

5' GGG CCC TGT CTC AGC TGG GGA

5' TGG CCT GGC TGC CCT GAG CAG

Temp.: 94oC/63 oC/72 oC

Time: 30s/1min/2min

Cycles: 30.

PCR reactions were carried out in a 25 or 50 m l volume containing 5-50 ng genomic DNA template.Detection of the different alleles from PCR amplified products were carried out by agarose gel electrophoresis. The gel mixture contained 2,0% agarose in a 1xTAE buffer with ethidium bromide.

Results and discussion

After studying 215 unrelated individuals of YNZ22 locus, 12 different alleles and 49 different genotypes were distinguishable in that locus ranging in size 170-940 b.p. The highest frequencies observed three alleles 240, 310 and 380 b.p. (0.1372, 0.1884 and 0.3256) - Table 1.

For Ig-JH locus a total of 24 different genotypes and 11 alleles were observed in 200 individuals. Two alleles - 570 and 670 b.p., were found to be relatively common for the Lithuanian sample, with frequencies of 0,4825 and 0,2600, respectively (Table 2). Distribution of YNZ22 genotypes in a samples of 215 Lithuanians are presented in Table 3, of Ig-JH genotypes in a samples of 200 Lithuanians - in Table 4.

From a forensic point of view the YNZ22 and Ig-JH polymorphisms have been examined in paternity cases. Based on the allele frequencies, heterozygosity index, power of discrimination, chance exclusion and Essen-Moller value were calculated. The combined values show a high efficiency of forensic studies.

Table 1. YNZ22 allele frequencies from 215 Lithuanian individuals

Allele

Allele size

n

F

1

170

22

0,0512

2

240

59

0,1372

3

310

81

0,1884

4

380

140

0,3256

5

450

17

0,0395

6

520

12

0,0279

7

590

9

0,0209

8

660

10

0,0233

9

730

24

0,0558

10

800

34

0,0791

11

870

8

0,0186

12

940

14

0,0325

13

1010  

-

-

14

1080  

-

-

14(12)

 

430

1,0000

Table 2. Ig-JH allele frequencies from 200 Lithuanian individuals

Allele

Allele size

n

F

07

420

2

0,0050

08

470

47

0,1175

09

520

10

0,0250

10

570

193

0,4825

11

620

1

0,0025

12

670

104

0,2600

13

720

3

0,0075

14

770

-

-

15

820

1

0,0025

16

870

11

0,0275

17

920

25

0,0625

18

970

3

0,0075

19

1020  

-

-

13 (11)

 

400

1,0000

Table 3. Distribution of YNZ22 genotypes in a sample of 215 Lithuanians

Genotype

n

F

Genotype

n

F

170-170

1

0,005

310-940

2

0,009

170-240

2

0,009

380-380

29

0,135

170-310

4

0,019

380-450

3

0,014

170-380

7

0,033

380-520

4

0,019

170-450

2

0,009

380-590

4

0,019

170-520

1

0,005

380-660

3

0,014

170-590

1

0,005

380-730

4

0,019

170-800

2

0,009

380-800

9

0,042

170-940

1

0,005

380-870

3

0,014

240-240

10

0,047

380-940

2

0,009

240-310

9

0,042

450-450

2

0,009

240-380

18

0,084

450-730

2

0,009

240-450

1

0,005

450-870

1

0,005

240-590

3

0,014

450-940

1

0,005

240-660

3

0,014

520-590

1

0,005

240-730

1

0,005

520-660

1

0,005

240-800

2

0,009

520-730

1

0,005

310-310

11

0,051

520-940

1

0,008

310-380

25

0,116

730-730

2

0,009

310-450

3

0,014

730-800

6

0,028

310-520

3

0,014

730-870

1

0,005

310-660

3

0,014

730-940

4

0,019

310-730

1

0,005

800-800

3

0,014

310-800

6

0,028

800-940

3

0,014

310-870

3

0,014

 

 

 

49 genotypes

Homozygosity (n = 58) = 0,270

Heterozygosity (n = 157) = 0,730

Table 4. Distribution of Ig-JH genotypes in a sample of 200 Lithuanians

Genotype

n

F

420-620

1

0,005

470-920

1

0,005

470-470

3

0,015

470-570

21 

0,105

470-670

16  

0,080

470-720

1

0,005

470-870

1

0,005

470-920

2

0,010

520-570

5

0,025

520-670

5

0,025

570-570

49  

0,245

570-670

51  

0,255

570-720

1

0,005

570-870

6

0,030

570-920

9

0,045

570-970

2

0,010

670-670

11  

0,055

670-820

1

0,005

670-870

2

0,010

670-920

6

0,030

670-970

1

0,005

720-920

1

0,005

870-920

2

0,010

920-920

2

0,010

24 genotypes

Homozygosity (n = 65) = 0,325

Heterozygosity (n = 135) = 0,675

References

1. Budowle B., Charkraborty R., Giusti A. W., Am. J. Hum. Genet., 1991, vol. 48, p. 137-144.

2. Gecz J., Nucleic Acids Res., 1991, vol. 19, p. 5806.

3. Gene M., Huguet E., Int. J. Leg.Med., 1995, vol. 107, p. 222-224.

4. Pinheiro M, Pontes M., Forensic Science Int., 1996, vol. 79, p. 23-29.



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