CAJ | 학술논문

目的比较CAPN3基因在外周血白细胞和骨骼肌组织中的剪切变异,探讨用外周血白细胞CAPN3 mRNA进行基因诊断的可行性.方法抽提正常人外周血和骨骼肌组织中总RNA,通过逆转录-聚合酶链反应和DNA测序确定CAPN3基因的cDNA序列,比较外周血白细胞和骨骼肌组织中CAPN3cDNA序列.结果由骨骼肌组织抽提的RNA进行逆转录合成cDNA为CAPN3基因全长cDNA,包含全部24个外显子;而由外周血白细胞抽提的RNA进行逆转录合成cDNA为CAPN3基因非全长cDNA,包含23个外显子,缺失了第15外显子.结论人的CAPN3基因在骨骼肌和外周血自细胞中存在着不同的剪切方式,若用外周血抽提RNA进行CAPN3基因的编码序列分析时会漏检第15外显子的突变.这提示从cDNA水平分析CAPN3基因突变时应采用患者肌肉组织,而非外周血白细胞.
목적 비교CAPN3기인재외주혈백세포화골격기조직중적전절변이,탐토용외주혈백세포CAPN3 mRNA진행기인진단적가행성.방법 추제정상인외주혈화골격기조직중총RNA,통과역전록-취합매련반응화DNA측서학정CAPN3기인적cDNA서렬,비교외주혈백세포화골격기조직중CAPN3cDNA서렬.결과 유골격기조직추제적RNA진행역전록합성cDNA위CAPN3기인전장cDNA,포함전부24개외현자;이유외주혈백세포추제적RNA진행역전록합성cDNA위CAPN3기인비전장cDNA,포함23개외현자,결실료제15외현자.결론 인적CAPN3기인재골격기화외주혈자세포중존재착불동적전절방식,약용외주혈추제RNA진행CAPN3기인적편마서렬분석시회루검제15외현자적돌변.저제시종cDNA수평분석CAPN3기인돌변시응채용환자기육조직,이비외주혈백세포.
Objective To investigate the splice variants of the calpain 3 gene existing in human skeletal muscle tissue and white blood cells, and to explore the feasibility of gene diagnosis using CAPN3 mRNA extracted from peripheral leukocytes. Methods Total RNA was extracted from peripheral blood and skeletal muscle tissue in healthy individuals. CAPN3 cDNAs were determined by reverse transcriptase polymerase chain reaction and DNA sequencing. CAPN3 cDNAs from peripheral leukocytes were compared with sequences obtained from skeletal muscle tissue. Results RT-PCR and DNA sequencing showed that the CAPN3 cDNAs comprised 24 exons in human skeletal muscle tissue, while the number of exons was 23 in white blood cells. Exon 15 was spliced out in human white blood cells. Conclusion Splice variants exist in human skeletal muscle tissue and white blood cells. Gene diagnosis may omit the mutations of exon 15 using mRNA extracted from peripheral leukocytes. These findings suggest that mutation analysis of the CAPN3 cDNA should use skeletal muscle tissue as materials instead of peripheral blood.