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Novel FOXL2 mutations in two Chinese families with blepharophimosis-ptosis-epicanthus inversus syndrome.

Xue M, Zheng J, Zhou Q, Hejtmancik JF, Wang Y, Li S - BMC Med. Genet. (2015)

Bottom Line: The sequencing results were analyzed using DNAstar software.A novel FOXL2 heterozygous indel mutation c.675_690delinsT, including a 16-bp deletion and a 1-bp(T) insertion (p.Ala226_Ala230del), which would result in deletion of 5 alanine residues of a poly-alanine (poly-Ala) tract in the protein, was identified in all affected members of family A.Our results expand the spectrum of FOXL2 mutations and provide additional structure-function insights into the FOXL2 protein.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, the First Affiliated Hospital of Anhui Medical University, Hefei, China. flyxuemin@163.com.

ABSTRACT

Background: Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a rare autosomal dominant disease. Mutations in the forkhead box L2 (FOXL2) gene cause two types of BPES distinguished by the presence (type I) and absence (type II) of premature ovarian failure (POF). The purpose of this study was to identify possible mutations in FOXL2 in two Chinese families with BPES.

Methods: Two large autosomal dominant Chinese BPES families were enrolled in this study. Genomic DNA was obtained from the leukocytes in peripheral venous blood. Four overlapping sets of primers were used to amplify the entire coding region and nearby intron sequences of the FOXL2 gene for mutations detection using polymerase chain reaction (PCR) and sequencing analyses. The sequencing results were analyzed using DNAstar software.

Results: All patients of the two families demonstrated typical features of BPES type II, including small palpebral fissures, ptosis, telecanthus, and epicanthus inversus without female infertility (POF). A novel FOXL2 heterozygous indel mutation c.675_690delinsT, including a 16-bp deletion and a 1-bp(T) insertion (p.Ala226_Ala230del), which would result in deletion of 5 alanine residues of a poly-alanine (poly-Ala) tract in the protein, was identified in all affected members of family A. A novel heterozygous missense mutation (c.223C > T, p.Leu75Phe) was identified in family B.

Conclusions: Two novel FOXL2 mutations were identified in Chinese families with BPES. Our results expand the spectrum of FOXL2 mutations and provide additional structure-function insights into the FOXL2 protein.

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Related in: MedlinePlus

The PCR products amplified by primer 4 in the Family a. Patients’ PCR products revealed two fragments of 304 and 289 bp using 6 % agarose gel electrophoresis. Normal individual contained a single fragment of 304 bp. The right lane is DNA marker (100bp)
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Fig2: The PCR products amplified by primer 4 in the Family a. Patients’ PCR products revealed two fragments of 304 and 289 bp using 6 % agarose gel electrophoresis. Normal individual contained a single fragment of 304 bp. The right lane is DNA marker (100bp)

Mentions: Analysis using agarose gel electrophoresis revealed two fragments in the PCR products amplified by primer 4 in the Family A patients, while those of unaffected individuals contained a single fragment (Fig. 2). Because these PCR products could not be adequately sequenced directly, the PCR fragments were purified and cloned into the PMD18-T vector (Takara Biotechnology (Dalian) Co., Ltd). The constructs were subsequently transformed into competent DH5α Escherichia coli. The plasmids were extracted from the positive clones. The cloned fragments of two types were identified by 6 % agarose gel electrophoresis after PCR using the primers mentioned above. The plasmids were subsequently sequenced on an ABI-3730 DNA Analyzer (Takara Biotechnology (Dalian) Co., Ltd) using BcaBEST sequencing primers RV-M: 5’-GAGCGGATAACAATTTCACACAGG-3’ and M13-47: 5’-CGCCAGGGTTTTCCCAGTCACGAC-3’.Fig. 2


Novel FOXL2 mutations in two Chinese families with blepharophimosis-ptosis-epicanthus inversus syndrome.

Xue M, Zheng J, Zhou Q, Hejtmancik JF, Wang Y, Li S - BMC Med. Genet. (2015)

The PCR products amplified by primer 4 in the Family a. Patients’ PCR products revealed two fragments of 304 and 289 bp using 6 % agarose gel electrophoresis. Normal individual contained a single fragment of 304 bp. The right lane is DNA marker (100bp)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4593235&req=5

Fig2: The PCR products amplified by primer 4 in the Family a. Patients’ PCR products revealed two fragments of 304 and 289 bp using 6 % agarose gel electrophoresis. Normal individual contained a single fragment of 304 bp. The right lane is DNA marker (100bp)
Mentions: Analysis using agarose gel electrophoresis revealed two fragments in the PCR products amplified by primer 4 in the Family A patients, while those of unaffected individuals contained a single fragment (Fig. 2). Because these PCR products could not be adequately sequenced directly, the PCR fragments were purified and cloned into the PMD18-T vector (Takara Biotechnology (Dalian) Co., Ltd). The constructs were subsequently transformed into competent DH5α Escherichia coli. The plasmids were extracted from the positive clones. The cloned fragments of two types were identified by 6 % agarose gel electrophoresis after PCR using the primers mentioned above. The plasmids were subsequently sequenced on an ABI-3730 DNA Analyzer (Takara Biotechnology (Dalian) Co., Ltd) using BcaBEST sequencing primers RV-M: 5’-GAGCGGATAACAATTTCACACAGG-3’ and M13-47: 5’-CGCCAGGGTTTTCCCAGTCACGAC-3’.Fig. 2

Bottom Line: The sequencing results were analyzed using DNAstar software.A novel FOXL2 heterozygous indel mutation c.675_690delinsT, including a 16-bp deletion and a 1-bp(T) insertion (p.Ala226_Ala230del), which would result in deletion of 5 alanine residues of a poly-alanine (poly-Ala) tract in the protein, was identified in all affected members of family A.Our results expand the spectrum of FOXL2 mutations and provide additional structure-function insights into the FOXL2 protein.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, the First Affiliated Hospital of Anhui Medical University, Hefei, China. flyxuemin@163.com.

ABSTRACT

Background: Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a rare autosomal dominant disease. Mutations in the forkhead box L2 (FOXL2) gene cause two types of BPES distinguished by the presence (type I) and absence (type II) of premature ovarian failure (POF). The purpose of this study was to identify possible mutations in FOXL2 in two Chinese families with BPES.

Methods: Two large autosomal dominant Chinese BPES families were enrolled in this study. Genomic DNA was obtained from the leukocytes in peripheral venous blood. Four overlapping sets of primers were used to amplify the entire coding region and nearby intron sequences of the FOXL2 gene for mutations detection using polymerase chain reaction (PCR) and sequencing analyses. The sequencing results were analyzed using DNAstar software.

Results: All patients of the two families demonstrated typical features of BPES type II, including small palpebral fissures, ptosis, telecanthus, and epicanthus inversus without female infertility (POF). A novel FOXL2 heterozygous indel mutation c.675_690delinsT, including a 16-bp deletion and a 1-bp(T) insertion (p.Ala226_Ala230del), which would result in deletion of 5 alanine residues of a poly-alanine (poly-Ala) tract in the protein, was identified in all affected members of family A. A novel heterozygous missense mutation (c.223C > T, p.Leu75Phe) was identified in family B.

Conclusions: Two novel FOXL2 mutations were identified in Chinese families with BPES. Our results expand the spectrum of FOXL2 mutations and provide additional structure-function insights into the FOXL2 protein.

Show MeSH
Related in: MedlinePlus