OBJECTIVES: To study the status of cochlear mitochondrial DNA (mtDNA) and to determine the location of mtDNA deletion in aged mice. METHODS: We detected cochlear mtDNA in 2, 7 - 10 and 17 - 19 month old mice by nested...OBJECTIVES: To study the status of cochlear mitochondrial DNA (mtDNA) and to determine the location of mtDNA deletion in aged mice. METHODS: We detected cochlear mtDNA in 2, 7 - 10 and 17 - 19 month old mice by nested polymerase chain reaction (PCR) and DNA sequencing. RESULTS: mtDNA3867bp deletions were found in the cochleae of aged mice. The deletion occurred within nt9103-nt12970 and were flanked by 15 base pair direct repeats. Comparing the incidence of mtDNA3867bp deletions, 17 - 19 month old mice (7/8) were significantly higher than 7 - 10 month old mice (4/16). The deletion was not observed in 2 month old mice (0/7). The ratio of deleted mtDNA/total mtDNA in 17 - 19 month old mice was higher than in 7 - 10 month old mice (P展开更多
根据NCBI上公布的弓形虫RH株的529bp基因序列(登录号:AF146527)用Primer Premier 5.0设计3条特异性引物TX1、TX2和TX3,建立土壤中弓形虫的半套式PCR检测方法,并将引物TX1与TX3扩增产物克隆到pMD18-T载体上进行测序.结果表明:该方法能扩...根据NCBI上公布的弓形虫RH株的529bp基因序列(登录号:AF146527)用Primer Premier 5.0设计3条特异性引物TX1、TX2和TX3,建立土壤中弓形虫的半套式PCR检测方法,并将引物TX1与TX3扩增产物克隆到pMD18-T载体上进行测序.结果表明:该方法能扩增出约350bp的片段,TX1与TX3扩增片段约520bp,克隆到pMD18-T载体上的521bp片段与所选取的弓形虫RH株的目的基因序列相似性100%.该检测方法特异性强,以犬新孢子虫、牛微小隐孢子虫、牛巴贝斯虫、环形泰勒虫、牛链球菌、猪链球菌、蒙氏肠球菌基因组为模板均未扩增出条带;灵敏性高,检测极限为20fg弓形虫DNA;在22份土壤样品中检出3份阳性样品.展开更多
基金ThisworkwassupportedbygrantsfromtheNationalOutstandingYouthSciencesFoundationofChina (No 39725026)andthePostdoctoralSciencesFoundationofChina (No [2 0 0 0 ]23)
文摘OBJECTIVES: To study the status of cochlear mitochondrial DNA (mtDNA) and to determine the location of mtDNA deletion in aged mice. METHODS: We detected cochlear mtDNA in 2, 7 - 10 and 17 - 19 month old mice by nested polymerase chain reaction (PCR) and DNA sequencing. RESULTS: mtDNA3867bp deletions were found in the cochleae of aged mice. The deletion occurred within nt9103-nt12970 and were flanked by 15 base pair direct repeats. Comparing the incidence of mtDNA3867bp deletions, 17 - 19 month old mice (7/8) were significantly higher than 7 - 10 month old mice (4/16). The deletion was not observed in 2 month old mice (0/7). The ratio of deleted mtDNA/total mtDNA in 17 - 19 month old mice was higher than in 7 - 10 month old mice (P
文摘根据NCBI上公布的弓形虫RH株的529bp基因序列(登录号:AF146527)用Primer Premier 5.0设计3条特异性引物TX1、TX2和TX3,建立土壤中弓形虫的半套式PCR检测方法,并将引物TX1与TX3扩增产物克隆到pMD18-T载体上进行测序.结果表明:该方法能扩增出约350bp的片段,TX1与TX3扩增片段约520bp,克隆到pMD18-T载体上的521bp片段与所选取的弓形虫RH株的目的基因序列相似性100%.该检测方法特异性强,以犬新孢子虫、牛微小隐孢子虫、牛巴贝斯虫、环形泰勒虫、牛链球菌、猪链球菌、蒙氏肠球菌基因组为模板均未扩增出条带;灵敏性高,检测极限为20fg弓形虫DNA;在22份土壤样品中检出3份阳性样品.
