Intron mutations of GABRG2 gene in patients with epilepsy combined with febrile seizures plus:an in vitro splicing assay

VernacularTitle:癫痫伴热性惊厥附加症患者GABRG2基因内含子突变的体外剪接分析
Author: Chao LIU ¹ ; Bin QIN ; Mei-Juan YU ; Yi-Wu SHI ; Wei-Ping LIAO
Author Information

1. 广州医学院附属第二医院
Keywords: Epilepsy; DNA mutation; GABRG2 gene; Splicing
From: Chinese Journal of Neuromedicine 2011;10(6):602-604
CountryChina
Language:Chinese
Abstract: Objective To screen the GABRG2 gene in Chinese patients diagnosed as having epilepsy with febrile seizures plus (EFS+) and analyze the in vitro splicing of intron mutations of GABRG2 gene. Methods After collecting the blood samples from patients with EFS+, all 9 coding exons and introns relevant to mRNA splice of GA BRG2 gene were sequenced by PCR. PCR products of exons 7, 8 and 9 and part of the introns of both ends of GABRG2 gene were cloned into the pTARGET vector to construct pTARGET-Exon-7-8-9 minigene vector and its Exon8+45C>T mutation vector.Wild-type and Exon8+45C>T mutation vector were transfected into HEK 293 cells and extracted RNA for RT-PCR. Results We did not detect mutation in GABRG2 gene coding region, but found 1 mutation in intron Exon8+45C>T. After splicing, the size of RT-PCR products of Wild-type and Exon8+45 OT mutation were both 522 bp. Conclusion Mutations in GABRG2 gene coding region are not likely to be substantially involved in the etiology of EFS+. Exon8+45C>T mutation does not affect the splicing of GABRG2 gene.