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981.
982.
983.
应用DPO-PCR方法特异性检测志贺氏菌 总被引:2,自引:0,他引:2
In this study, a dual-priming oligonucleotide (DPO)-based PCR method for specific detection of Shigella was established using ipaH gene of Shigella as the target gene. The results showed that detection sensitivity of the DPO-PCR method was 1.65×102 CFU/mL. Compared with conventional PCR primers, the DPO primers were easily designed, which simplified the design procedure of PCR primers. DPO primers were not sensitive to annealing temperature, which could efficiently amplify the target gene in the annealing temperature range from 50 to 70 ℃. Moreover, due to the special structure of DPO primers, it had a higher specificity than conventional PCR primers, and none nonspecific amplifications were produced in reaction. In the practical application, tests on 133 samples including frozen/fresh meat, fruits and vegetables, fresh milk, eggs and cooked food by the DPO-PCR method showed that 15 samples were Shigella positive, which were in accordance with the testing results using GB method (GB/T 4789.5-2012), showing good practicality and reliability. The DPO-PCR method provided a new tool for fast and accurate detection of Shigella. 相似文献
984.
将RT-PCR检测为猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)阳性的肺组织碾磨、过滤后接种MARC-145细胞,在细胞上连续传代,上清传至第二代后能产生细胞病变。通过RT-PCR方法可以检测到释放在细胞上清中PRRSV RNA。间接免疫荧光试验也能检测细胞内PRRSV N蛋白的表达,表明该PRRSV毒株成功地在MARC-145细胞中分离,命名为PRRSV NN1396株。对NN1396分离株GP5基因进行克隆测序分析,结果表明该毒株与NCBI登录的PRRSV GP5核苷酸同源性为62.9%~98.2%,与PRRSV原型毒株VR-2332的同源性为87.9%,与高致病性毒株PRRSV JXA1同源性为98.0%,通过GP5的遗传进化分析发现所分离的PRRSV为北美型PRRSV,且与高致病性PRRSV分布在同一个小分支上;对部分nsp2基因的测序、比对结果表明,该部分基因与NCBI登录的参考序列VR-2332氨基酸同源性为73.0%,与JXA1的同源性为94.8%。NN1396分离株Nsp2蛋白含有高致病性PRRSV特异的1+29个氨基酸的缺失,此外,在缺失29个氨基酸区域的上游,含有连续19个氨基酸缺失。该PRRSV毒株的分离和Nsp2蛋白部分序列的缺失鉴定,为下一步研究Nsp2部分氨基酸缺失对病毒生物学特性影响奠定基础。 相似文献
985.
核因子-κB(NF-κB)在炎症反应中起着关键的促进作用,并且受其他多种因素的调节。Nfkbiz基因编码的IκB?蛋白是IκB家族成员之一,在炎症反应中起着调节NF-κB的作用。本研究采用荧光实时定量PCR分析Nfkbiz基因的表达情况,发现在p53特定刺激源作用下Nfkbiz基因mRNA丰度显著降低。分析Nfkbiz基因序列,发现其基因中存在可能的p53反应元件,通过构建荧光素酶报告质粒及双荧光素酶报告试验和染色质免疫共沉淀试验,证明Nfkbiz基因中含有p53反应元件并且受p53蛋白调节。以上结果初步证明Nfkbiz是受p53蛋白负向调控的靶基因。 相似文献
986.
987.
988.
989.
- 1. Baiji were sighted 17 times during three recent simultaneous multi‐vessel surveys in the Yangtze River, China (November 4–10, 1997; December 4–9, 1998; October 31–November 5, 1999). There were 11 sightings in 1997 (consisting of 17 animals), five in 1998 (seven animals), and two in 1999 (four animals). It was concluded that 13 individuals could be considered as a minimum number of the baiji currently in the Yangtze River.
- 2. An annual rate of population decrease was roughly estimated as 10%. From the body sizes observed, the proportions of old, adult and immature individuals were approximately estimated at 57, 26, and 17% respectively.
- 3. Baiji showed a significant attraction to confluences and sand bars with large eddies. The present distribution range of the baiji is less than 1400 km in length in the Yangtze main river. Distances between the two nearest groups of baiji appear to be increasing.
- 4. Two typical sightings are described, in which surfacing and movements of baiji were recorded. Baiji were often found swimming together with finless porpoise. In the surveys they occurred in the same group in 63% of occurrences. Interactions between baiji and finless porpoise are described and discussed.
- 5. Human activities are the main threats to the baiji. Illegal electrical fishing accounted for 40% of known mortalities during the 1990s. Engineering explosions for maintaining navigation channels have become another main cause of baiji deaths. The last hope of saving the species may be to translocate the remaining baiji into a semi‐captive reserve, known as the ‘Baiji Semi‐natural Reserve’.
990.
运用地统计学和GIS相结合对琼中县橡胶园土壤磷素营养进行了空间变异特征及分区管理研究。结果表明,琼中县胶园各土层土壤全磷和有效磷含量的最大相关距离是17.23~52.09 km,全磷含量以大块状变异为主,有效磷含量各等级的分布为大、小块状交叉分布;琼中县橡胶园0~20 cm土层、20~40 cm土层全磷含量处于第五等级的极低水平的比例分别为91.22%和82.8%,0~20 cm土层有效磷含量多数处于中等偏上水平,而20~40 cm土层有效磷含量处于第5等级及以下的极低水平的比例为60.6%;应用空间叠加方法,进行了橡胶园土壤磷素管理分区,琼中县胶园可分为11个管理分区,在胶园施肥管理中应根据不同管理分区的特点采取不同的磷素营养管理措施。 相似文献