Pan Yu-Chun, Wu Wei-Qing, Xie Jian-Sheng, Luo Cai-Qun, Hao Ying
Prenatal Diagnosis Center, Shenzhen Maternity and Child Health Care Hospital, Southern Medical University, Shenzhen, Guangdong 518000, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2017 Mar;19(3):308-312. doi: 10.7499/j.issn.1008-8830.2017.03.011.
High-throughput sequencing was performed for the peripheral blood DNA from two probands in the family with tuberous sclerosis complex (TSC) to determine the sequences of TSC-related genes TSC1 and TSC2 and their splicing regions and identify mutation sites. Amplification primers were designed for the mutation sites and polymerase chain reaction and Sanger sequencing were used to verify the sequences of peripheral blood DNA from the probands and their parents. The two probands had c.3981-3982 insA (p.Asp1327AspfsX87) and c.4013-4014 delCA (p.Ser1338Cysfs) heterozygous mutations, respectively, in the TSC2 gene. The parents of proband 1 had no abnormalities at these two loci; the mother of proband 2 had c.4013-4014 delCA heterozygous mutation in the TSC2 gene, while the father and the grandparents of proband 2 had no abnormalities. c.3981-3982 insA mutation may cause early coding termination of amino acid sequence at the 1413th site, and c.4013-4014 delCA mutation may cause early coding termination of amino acid sequence at the 1412th site. These two mutations are the pathogenic mutations for families 1 and 2, respectively, and both of them are novel frameshift mutations, but their association with the disease needs to be further verified by mutant protein function cell model and animal model.
对结节性硬化症(TSC)家系中的两名先证者的外周血DNA进行高通量测序,以确定TSC相关基因TSC1和TSC2及其剪接区域的序列,并鉴定突变位点。针对突变位点设计扩增引物,采用聚合酶链反应和桑格测序法验证先证者及其父母外周血DNA的序列。两名先证者的TSC2基因分别有c.3981-3982 insA(p.Asp1327AspfsX87)和c.4013-4014 delCA(p.Ser1338Cysfs)杂合突变。先证者1的父母在这两个位点无异常;先证者2的母亲在TSC2基因中有c.4013-4014 delCA杂合突变,而先证者2的父亲和祖父母无异常。c.3981-3982 insA突变可能导致氨基酸序列在第1413位提前编码终止,c.4013-4014 delCA突变可能导致氨基酸序列在第1412位提前编码终止。这两个突变分别是家系1和家系2的致病突变,且均为新的移码突变,但其与疾病的关联还需通过突变蛋白功能细胞模型和动物模型进一步验证。