植物内生菌16S rRNA基因扩增子高通量测序中降低宿主污染的方法

植物内生菌能够帮助植物生长,增强其抗逆性与抗病性,研究植物内生菌对了解植物入侵机制、保护生态环境与农业的健康发展具有重要意义.通过阅读相关文献,总结归纳了降低植物内生菌16SrRNA基因扩增子高通量测序(16S-seq)中宿主基因污染的4种方法,即:①寻找特异性错配引物用来扩增细菌16S rRNA基因;②添加特异性阻断...     

生物硝化池污水中硝化细菌的快速定量研究

实验采用聚合酶链式反应(PCR)技术与最大几率数法(MPN)相结合的MPNPCR法对生物硝化池污水中的硝化细菌进行快速定量.所用的一对PCR引物是在对硝化细菌的16SrRNA基因进行系统比较的基础上设计合成的, 可以扩增出大小为388bp的DNA片段.以从生物硝化池污水中抽提的含硝化细菌DNA的混合DNA为模板, 进行PCR扩增并确定合适的扩增条件.运用MPNPCR法进行定量检测的整个过程可在几小时之内完成.     

Normal pregnancy after preimplantation DNA diagnosis of a dystrophin gene deletion

To perform preimplantation DNA diagnosis for Duchenne muscular dystrophy (DMD) in a female carrier of a dystrophin gene deletion of exons 3–18, we developed a polymerase chain reaction (PCR)-based assay of exon 17 sequences. Exon 17 was efficiently amplified in all 50 single blastomeres of normal control embryos and in five blastomeres of one male embryo of the DMD carrier obtained after a first preimplantation diagnosis (PID) for gender determination. In ten blastomeres of another two male embryos of the DMD carrier, no PCR signals were observed, probably as a result of the deletion. After intracytoplasmic sperm injection, embryos were analysed for exon 17 and three of the four embryos showing normal PCR signals were replaced, resulting in a singleton pregnancy. Prenatal diagnosis showed a female karyotype and DNA analysis indicated that the fetus was not a DMD carrier.     

长期施用四环素残留猪粪对土壤中耐药菌及抗性基因形成的影响

探讨了长期施用含兽用四环素药物残留的畜禽粪便对土壤环境中耐药菌及抗性基因形成的影响.分秋季和夏季两次采集苏北黄淮地区沭阳市某养猪场周边长期用含有四环素残留的猪粪作为肥料的耕地土壤,并采集当地没有施加过猪粪的耕地土壤作为对照.分析样品中耐药菌的组成,同时利用PCR技术研究3种目前常见的四环素抗性基因(tetA、tetC、tetE).结果表明,从秋季土壤样品中共分离出59株耐药菌,属于13个菌属,夏季土壤中共分离出35株耐药菌,属于10个菌属,其中致病菌数占总耐药菌数的比例高达38.14%,而对照组中的3株耐药菌只属于一个菌属(Streptomyces).PCR结果显示所有的耐药菌上都携带了抗性基因,tetC是含量最高的抗性基因.施用过含有四环素残留猪粪的土壤样品中四环素的残留含量为41.1~61.9μg·kg-1,干土中四环素抗性基因含量为4.63×105~37.42×105copies·g-1,通过将土壤中四环素的残留量与土壤中抗性基因的量进行拟合,结果发现在一定范围内,四环素残留量与四环素抗性基因量存在着显著的正相关关系.研究还发现合适的气候条件对耐药菌和抗性基因的形成有较好的促进作用.     

水体中抗生素的检测及去除方法研究综述

抗生素是目前世界上应用最广泛的药物之一。大量抗生素的使用,不可避免地给自然环境造成巨大的压力。由于污水截流不彻底等原因,工业、养殖及医疗活动等进入环境中的抗生素通过各种途径进入饮用水体,对水质安全及人类健康构成威胁。针对抗生素对水环境造成的污染,讨论目前国内外水环境中抗生素检测技术和去除手段的研究情况,主要对水中抗生素的富集方法、仪器检测及臭氧氧化去除技术进行分析,最后对抗生素污染的研究方向进行了展望。     

环境样品中甲基汞的分析方法综述

甲基汞在环境中分布广泛,可以对人类的生活产生威胁,故对甲基汞的研究具有重要意义。但是环境样品中甲基汞的含量通常很低,一般为10-9级,甚至到10-12级,非常难于测定。文中详细论述和比较了各种常见环境样品中的预处理和分析方法,如沉积物、土壤、生物样、水样、大气等,并讨论了各种方法的优缺点。     

Prenatal diagnosis of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency in a family with a previous fatal case of sudden unexpected death in childhood

Medium-chain acyl-CoA dehydrogenase (MCAD) deficiency is a potentially fatal inherited disease with a carrier frequency of approximately 1:100 in most Caucasian populations. The disease is implicated in sudden unexpected death in childhood. A prevalent disease-causing point mutation (A985G) in the MCAD gene has been characterized, thus rendering diagnosis easy in the majority of cases. Since the clinical spectrum of MCAD deficiency ranges from death in the first days of life to an asymptomatic life, there are probably other genetic factors—in addition to MCAD mutations—involved in the expression of the disease. Thus, families who have experienced the death of a child from MCAD deficiency might have an increased risk of a seriously affected subsequent child. In such a family we have therefore performed a prenatal diagnosis on a chorionic villus sample by a highly specific and sensitive polymerase chain reaction (PCR) assay for the G985 mutation. The analysis was positive and resulted in abortion. We verified the diagnosis by direct analysis on blood spots and other tissue material from the aborted fetus and from family members.