32 6 Vol. 32 No. 6 2011 12 Journal of Jinan University Medicine Edition Dec. 2011 Ⅰ TYR 1 1 1 2 1. 510632 2. 510080 1 OCA TYR PCR TYR HA DHPLC DNA TYR c. 232 _ 233insGGG /c. 841G > T c. 232_233insGGG c. 841G > T E281X c. 232_233insGGG c. 841G > T HA DHPLC c. 841G > T Ⅰ TYR R394. 11 A 1000-9965 2011 06-0624 - 05 Application of the mutation analysis for oculocutaneous albinism typeⅠ in the prenatal diagnosis and a novel pathogenic mutation of TYR gene MIAO Chun-yue 1 WEN Ren-qing 1 ZHENG Hui 1 LI Hong-yi 2 1. Department of Physiology Medical College Jinan University Guangzhou 510632 China 2. Department of Medical Genetics Zhongshan Medical College of Sun Yat-Sen University Guangzhou 510080 China Abstract Aim Mutation analysis and prenatal diagnosis for the mutation of tyrosinase TYR gene in a family with oculocutaneous albinism type Ⅰ OCA1. Methods Polymerase chain reaction PCR technique was applied to amplify the regions of exon exon-intron and promoter of TYR gene while heteroduplex analysis HA denaturing high performance liquid chromatography DHPLC technique and DNA sequencing technology were used to analyze gene mutation sites of the proband and her parents. The pathogenic mutations were identified and the fetus genotypes were obtained by detecting TYR gene corresponding sites of the DNA sequence. Results The patient of this family was found to be a compound heterozygote with mutants c. 232_233insGGG and c. 841G > T. The parents of proband showed the same mutants c. 232_233insGGG and c. 841G > T. The nonsense mutation E281X was found to be pathogenic. However the fetus got neither of the two mutations. The results obtained by HA and DHPLC methods were fully consistent with the DNA sequencing results. Conclusion The proband's genotype was deter- 2011-07 - 05 30672003 A2009351 1984 - Tel 020-85220260 E-mail tlihzh@ jnu. edu. cn
6 Ⅰ TYR 625 mined which was found to be a novel pathogenic mutation of TYR gene c. 841G > T. The fetus did not inherit any pathogenic mutations. Key words oculocutaneous albinism type 1 TYR gene gene mutation prenatal diagnosis Albinism 100 200 DNA 2 DNA Taq oculocutaneous albinism OCA 1 /20 000OCA 100 bp DNA OCA1 ~ OCA4 4 1-4 OCA1 tyrosinase TYR PCR OCA PerkinElmer WAVE 2100 1 5-10 Transgenomic uvp-gds7600 11 1. 2 1 DNA HA DHPLC TYR 1-7 DNA TYR OCA1 OCA1A OCA1B OCA1A OCA1B DNA 2 ml EDTA 6-9 20 OCA DNA 2 PCR PCR UCSC Genome Bioinformatics 12 TYR OCA1 1 NM-000372 1 1. 1 200 bp DNA TYR 5 - PCR 30 μl Mg 2 + 2. 5 1 OCA mmol /L dntp 2. 0 mmol /L Taq 1. 5 U 4 0. 17 μmol /L DNA 0. 5 μg 10 3 μl 94 3 min 94 45 ~ 60 s 49 ~ 65 OCA1 30 ~ 45 s 72 45 ~ 60 s 35 72 10 min 4 TYR 2%
626 32 3 PCR 8% 0. 5 TBE 2 300 V 30 min PCR 18 μl 2 μl 200 V 3. 5 h 0. 28 mol /L + 10% 4 PCR 95 WAVE DNA DHPLC WAVEMARKTM DNA ± 2 1 1 1 2 3 4 5 DNA TYR. DNA 2 2 DHPLC 2 DNA PCR 2 _233insGGG /c. 841G > T TYR 2. 1 PCR c. 232_233insGGG c. 841G > T DNA TYR TYR 2 % c. 232_233insGGG c. 841G > T 3 2. 2 DNA 2. 3 TYR 1 2 c. 841G > T 1 100 3 c. 841G > Tc. 841G / > T E281X 2 1 1 1 2 3 4. c. 232 3 c. 232 _ 233insGGG /c. 841G > T TYR c. 232 _ 233insGGG c. 841G > T OCA1 c. 841G > T 100 13 c. 1 1 841G TYR 281 281 DHPLC CuA CuB E 2 DHPLC
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