title: [Cloning and sequence analysis of SLC25A13 transcripts in human amniocytes].
abstract: OBJECTIVE
  This research intends to amplify the entire coding region sequences of SLC25A13 mRNA which encodes citrin, and to investigate sequence features of the transcripts for this gene in cultured human amniocytes. This study will provide laboratory evidence for prenatal diagnosis of neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) at mRNA level.
  
  
  METHODS
  One amniocyte sample was collected from a pregnant woman who underwent prenatal diagnosis of citrin deficiency and whose fetus has proven a carrier of 851del4 mutation by genomic DNA analysis. Another amniocyte sample, as a control, was from a fetus without family history of citrin deficiency. Total RNA was extracted from cultured amniocytes, cDNA was synthesized, and then nested-PCR was performed to amplify the entire coding region sequences of SLC25A13. The PCR products were cloned and analyzed by sequencing.
  
  
  RESULTS
  The entire coding region of SLC25A13 gene was successful amplified from two cultured human amniocytes. The splice variant of SLC25A13, SLCA (normal mRNA), was identified in the two samples. SLCB (CAG insertion between exon 9-10) was identified in the control. SLCC (exon 5-11 skipping), but not transcriptional product from the allele with 851del4 mutation, was identified in the 851del4 mutation carrier.
  
  
  CONCLUSIONS
  This study demonstrated that the entire coding region of SLC25A13 cDNA can be successfully amplified from two cultured human amniocytes, and revealed exon 5-11 skipping as a novel SLC25A13 transcript. Normal mRNA predominated in the transcripts in normal control and 851del4 mutation carrier, suggesting that the two fetuses were not at risk for NICCD. These SLC25A13 transcription features provided laboratory evidence for prenatal diagnosis of NICCD.
authors.affiliation: Department of Pediatrics, First Affiliated Hospital, Jinan University, Guangzhou 510630, China.
authors.forename: Zhan-Hui
authors.initials: ZH
authors.lastname: Zhang
authors.forename: Xin-Jing
authors.initials: XJ
authors.lastname: Zhao
authors.forename: Yuan-Zong
authors.initials: YZ
authors.lastname: Song
authors.forename: Xiao-Mei
authors.initials: XM
authors.lastname: Tang
authors.forename: Qing-Bing
authors.initials: QB
authors.lastname: Zha
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology* / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000172:deficiency; D002780:Cholestasis, Intrahepatic / Q000175:diagnosis; D003001:Cloning, Molecular; D005260:Female; D006801:Humans; D033681:Mitochondrial Membrane Transport Proteins / Q000235:genetics*; D027361:Organic Anion Transporters / Q000172:deficiency; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis*; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
chemical_list: D002135:Calcium-Binding Proteins; D033681:Mitochondrial Membrane Transport Proteins; D027361:Organic Anion Transporters; D012333:RNA, Messenger; C119629:SLC25A13 protein, human; C008822:citrin