title: Cloning and sequence analysis of SLC25A13 transcripts in human amniocytes.
abstract: OBJECTIVE
  To amplify the entire ORF of SLC25A13 cDNA which encodes citrin, a liver-type mitochondrial aspartate-glutamate carrier, and to investigate sequence feature of the transcripts for this gene in cultured human amniocytes. This study will provide laboratory evidences for prenatal diagnosis of NICCD at mRNA level.
  
  
  METHODS
  Total RNA was extracted from cultured amniocytes, and cDNA was synthesized, and then nest PCR was performed to amplify the entire ORF sequences of SLC25A13. The PCR products were purified, cloned, sequenced, and aligned with the genomic DNA of SLC25A13 to analyze the alternative splicing pattern.
  
  
  RESULTS
  The entire ORF of SLC25A13 gene was successfully amplified. Three splice variants of SLC25A13, i.e., SLCA (normal mRNA), SLCB (CAG insertion between exon 9-10) and SLCC (exon 5-11 skipping), were identified in the subjects. However, no abnormal mRNA from the allele with mutation 851del4 was detected in the amniocytes cultured from a carrier fetus of this mutation.
  
  
  CONCLUSIONS
  This study demonstrated that the entire ORF of SLC25A13 cDNA can be successfully amplified from cultured human amniocytes, and revealed exon 5-11 skipping as a novel SLC25A13 transcript. Normal mRNA occupied majority of the transcripts in normal control and heterozygous amniocytes which contained normal allele and 851del4 mutation, indicating that the two fetuses wouldn't suffer from NICCD. These SLC25A13 transcription features provided laboratory evidences for prenatal diagnosis of NICCD.
authors.affiliation: 1 Department of Pediatrics, 2 Department of Obstetrics and Gynecology, 3 Department of Fetal Medicine, the First Affiliated Hospital, Jinan University, Guangzhou 510630, China.
authors.forename: Zhan-Hui
authors.initials: ZH
authors.lastname: Zhang
authors.affiliation: 1 Department of Pediatrics, 2 Department of Obstetrics and Gynecology, 3 Department of Fetal Medicine, the First Affiliated Hospital, Jinan University, Guangzhou 510630, China.
authors.forename: Xin-Jing
authors.initials: XJ
authors.lastname: Zhao
authors.affiliation: 1 Department of Pediatrics, 2 Department of Obstetrics and Gynecology, 3 Department of Fetal Medicine, the First Affiliated Hospital, Jinan University, Guangzhou 510630, China.
authors.forename: Yuan-Zong
authors.initials: YZ
authors.lastname: Song
authors.affiliation: 1 Department of Pediatrics, 2 Department of Obstetrics and Gynecology, 3 Department of Fetal Medicine, the First Affiliated Hospital, Jinan University, Guangzhou 510630, China.
authors.forename: Xiao-Mei
authors.initials: XM
authors.lastname: Tang
authors.affiliation: 1 Department of Pediatrics, 2 Department of Obstetrics and Gynecology, 3 Department of Fetal Medicine, the First Affiliated Hospital, Jinan University, Guangzhou 510630, China.
authors.forename: Qing-Bing
authors.initials: QB
authors.lastname: Zha
keywords: SLC25A13 gene; alternative splicing; amniocyte; nest PCR
references.citation: J Inherit Metab Dis. 2007 Apr;30(2):139-44
references.pmid: 17323144
references.citation: Zhongguo Dang Dai Er Ke Za Zhi. 2009 May;11(5):328-32
references.pmid: 19470249
references.citation: J Hum Genet. 2008;53(6):534-45
references.pmid: 18392553
references.citation: Hum Mol Genet. 1999;8(10):1893-900
references.pmid: 10469842
references.citation: Zhongguo Dang Dai Er Ke Za Zhi. 2006 Apr;8(2):125-8
references.pmid: 16613706
references.citation: Tohoku J Exp Med. 2011;225(4):273-6
references.pmid: 22095253
references.citation: Nat Rev Mol Cell Biol. 2004 Feb;5(2):89-99
references.pmid: 15040442
references.citation: Clin Biochem. 2008 May;41(7-8):616-20
references.pmid: 18291102
references.citation: Mol Genet Metab. 2007 Jan;90(1):30-6
references.pmid: 17092749
references.citation: J Hum Genet. 2005;50(7):338-46
references.pmid: 16059747
references.citation: Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2010 Apr;27(2):180-5
references.pmid: 20376801
references.citation: Pediatr Transplant. 2010 Nov;14(7):E86-8
references.pmid: 19413723
references.citation: Zhongguo Dang Dai Er Ke Za Zhi. 2011 Apr;13(4):303-8
references.pmid: 21507300
references.citation: Zhonghua Er Ke Za Zhi. 2009 Aug;47(8):624-7
references.pmid: 19951499
references.citation: Eur J Cell Biol. 2011 Jun-Jul;90(6-7):603-11
references.pmid: 21324547
references.citation: Curr Diab Rep. 2010 Dec;10(6):444-51
references.pmid: 20878273
references.citation: Nature. 2008 Nov 27;456(7221):470-6
references.pmid: 18978772
references.citation: J Hum Genet. 2002;47(7):333-41
references.pmid: 12111366
references.citation: Mol Genet Metab. 2009 Jan;96(1):44-9
references.pmid: 19036621
references.citation: Dig Liver Dis. 2009 Sep;41(9):683-9
references.pmid: 19185551
references.citation: Int J Mol Med. 2011 Jul;28(1):33-40
references.pmid: 21424115
references.citation: Nat Genet. 1999 Jun;22(2):159-63
references.pmid: 10369257
references.citation: Biochemistry. 2000 Feb 22;39(7):1807-16
references.pmid: 10677231