中国当代儿科杂志2012，Vol.14Issue(3)：221-225,5.
人羊水细胞中SLC25A13基因转录产物的克隆和序列分析
Cloning and sequence analysis of SLC25A13 transcripts in human amniocytes
摘要
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 85]del4 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.关键词
SLC25A13基因/巢式PCR/Citrin缺陷导致的新生儿肝内胆汁淤积症/羊水细胞Key words
SLC25A13/ Nest PCR/ Neonatal intrahepatic cholestasis caused by citrin deficiency/ Amniocyte分类
医药卫生引用本文复制引用
张占会,赵新景,宋元宗,汤小湄,查庆兵..人羊水细胞中SLC25A13基因转录产物的克隆和序列分析[J].中国当代儿科杂志,2012,14(3):221-225,5.基金项目
国家自然科学基金资助项目(No.81070279). （No.81070279）
