Limits...
Nucleotide sequence analysis of NIPBL gene in Indian Cornelia de Lange syndrome cases.

Bajaj S, Ranade S, Gambhir P - Indian J Hum Genet (2013)

Bottom Line: The polymorphisms are in the region of intron 1 and in exon 10.The polymorphism C/A is present in intron 1 region and polymorphism T/G in exon 10.The intronic region polymorphism may have a role in intron splicing whereas the polymorphism in exon 10 results in amino acid change (Val to Gly).

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, University of Pune, Pune, Maharashtra, India.

ABSTRACT

Background: Cornelia de Lange syndrome (CdLS) is a multisystem developmental disorder in children. The disorder is caused mainly due to mutations in Nipped-B-like protein. The molecular data for CdLS is available from developed countries, but not available in developing countries like India. In the present study, the hotspot region of NIPBL gene was screened by Polymerase Chain Reaction which includes exon 2, 22, 42, and a biggest exon 10, in six CdLS patients and ten controls.

Materials and methods: The method adopted in present study was amplification of the target exon by using polymerase chain reaction, qualitative confirmation of amplicons by Agarose Gel Electrophoresis and use of amplicons for Conformation Sensitive Gel Electrophoresis to find heteroduplex formation followed by sequencing.

Results: We report two polymorphisms in the studied region of gene NIPBL. The polymorphisms are in the region of intron 1 and in exon 10. The polymorphism C/A is present in intron 1 region and polymorphism T/G in exon 10.

Conclusion: The intronic region polymorphism may have a role in intron splicing whereas the polymorphism in exon 10 results in amino acid change (Val to Gly). These polymorphisms are disease associated as these are found in CdLS patients only and not in controls.

No MeSH data available.


Related in: MedlinePlus

Well No 1-100 bp ladder, 2-7 amplified product exon 10 (A-F primers) well 8 and 9 - exon 2 and 42
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3722637&req=5

Figure 1: Well No 1-100 bp ladder, 2-7 amplified product exon 10 (A-F primers) well 8 and 9 - exon 2 and 42

Mentions: The patients studied were phenotypically shown the following features: Mental retardation, short stature, hypertrichosis, presence of microcephaly, synophrys, low antiriot hairline, narrow forehead, hoarse voice, marked on back, restricted movements of elbows, bilateral simian crease, short little finger with clinodactyly, hypoplastic nipples, shield chest, shawl scrotum, micropenis, and small testes. Family history for disease was nil. The families of these CdLS patients were studied for mutational analysis using PCR. The patients DNA isolated from leukocytes were subjected to amplification using primers for exons 2, 10, 22, and 42. Figure 1 shows the separation of amplicons on Agarose Gel Electrophoresis.


Nucleotide sequence analysis of NIPBL gene in Indian Cornelia de Lange syndrome cases.

Bajaj S, Ranade S, Gambhir P - Indian J Hum Genet (2013)

Well No 1-100 bp ladder, 2-7 amplified product exon 10 (A-F primers) well 8 and 9 - exon 2 and 42
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3722637&req=5

Figure 1: Well No 1-100 bp ladder, 2-7 amplified product exon 10 (A-F primers) well 8 and 9 - exon 2 and 42
Mentions: The patients studied were phenotypically shown the following features: Mental retardation, short stature, hypertrichosis, presence of microcephaly, synophrys, low antiriot hairline, narrow forehead, hoarse voice, marked on back, restricted movements of elbows, bilateral simian crease, short little finger with clinodactyly, hypoplastic nipples, shield chest, shawl scrotum, micropenis, and small testes. Family history for disease was nil. The families of these CdLS patients were studied for mutational analysis using PCR. The patients DNA isolated from leukocytes were subjected to amplification using primers for exons 2, 10, 22, and 42. Figure 1 shows the separation of amplicons on Agarose Gel Electrophoresis.

Bottom Line: The polymorphisms are in the region of intron 1 and in exon 10.The polymorphism C/A is present in intron 1 region and polymorphism T/G in exon 10.The intronic region polymorphism may have a role in intron splicing whereas the polymorphism in exon 10 results in amino acid change (Val to Gly).

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, University of Pune, Pune, Maharashtra, India.

ABSTRACT

Background: Cornelia de Lange syndrome (CdLS) is a multisystem developmental disorder in children. The disorder is caused mainly due to mutations in Nipped-B-like protein. The molecular data for CdLS is available from developed countries, but not available in developing countries like India. In the present study, the hotspot region of NIPBL gene was screened by Polymerase Chain Reaction which includes exon 2, 22, 42, and a biggest exon 10, in six CdLS patients and ten controls.

Materials and methods: The method adopted in present study was amplification of the target exon by using polymerase chain reaction, qualitative confirmation of amplicons by Agarose Gel Electrophoresis and use of amplicons for Conformation Sensitive Gel Electrophoresis to find heteroduplex formation followed by sequencing.

Results: We report two polymorphisms in the studied region of gene NIPBL. The polymorphisms are in the region of intron 1 and in exon 10. The polymorphism C/A is present in intron 1 region and polymorphism T/G in exon 10.

Conclusion: The intronic region polymorphism may have a role in intron splicing whereas the polymorphism in exon 10 results in amino acid change (Val to Gly). These polymorphisms are disease associated as these are found in CdLS patients only and not in controls.

No MeSH data available.


Related in: MedlinePlus