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121.
PCR及其衍生技术在基因突变检测中的应用 总被引:3,自引:1,他引:3
许多人类遗传性疾病及某些抗艾滋病药物的抗性乃至细菌对某些抗生素的抗药性通常源于基因突变。本文对近年来在基因突变检测中应用日益广泛的各种PCR 衍生技术作一综述;重点介绍了错配PCR技术,以及我们实验室近期报道的一种快速检测喹诺酮类药物耐药大肠杆菌的错配PCR方案。
Abstract:Many inherited diseases and drug resistance have been attributed to mutations in corresponding genes.In this paper,several techniques based on PCR used in diagnosis were concluded.The development and research progress of Mismatch PCR were discussed in details.Some information about an assay that we developed for detection of antimicrobial resistance to quinolones was also described. 相似文献
122.
123.
Miao Liu Yanfei Ru Yihua Gu Jianan Tang Tiancheng Zhang Jun Wu Fudong Yu Yao Yuan Chen Xu Jian Wang Huijuan Shi 《Biochimica et Biophysica Acta (BBA)/General Subjects》2018,1862(3):660-668
Background
We previously cloned the Ssp411 gene. We found that the Ssp411 protein is predominantly expressed in elongated spermatids in the rat testis in a stage-dependent manner. Although our findings strongly suggested that Ssp411 might play an important role in mammalian spermatogenesis, this hypothesis has not been studied.Methods
We first used real-time PCR, Western blotting and immunohistochemistry to confirm that the expression pattern of Ssp411 in several murine tissues is similar to its expression pattern in corresponding rat tissues. To better understand the roles of Ssp411 in male reproduction in vivo, we identified and characterized an Ssp411 expression-disrupted murine strain (Ssp411PB/PB) that was generated by piggyBac (PB) transposon insertion. We studied Ssp411-interacting proteins using proteome microarray, co-IP and GST pull-down assay.Results
Both Ssp411 mRNA and protein were detected exclusively in spermatids after step 9 during spermiogenesis in testis. Phenotypic analysis suggested that only Ssp411PB/PB males are sterile. These males have smaller testes, reduced sperm counts, decreased sperm motility and deformed spermatozoa. Microscopy analysis indicated that the manchette, a structurally reshaped sperm head, is aberrant in Ssp411PB/PB spermatids. The results of proteome microarray analysis and GST pull-down assays suggested that Ssp411 participates the ubiquitin-proteasome system by interacting with PSMC3. This has been reported to be manchette-associated and important for the head shaping of spermatids.Conclusions
Our study suggested that Ssp411 is required for spermiogenesis. It seems to play a role in sperm head shaping. The lack of Ssp411 causes sperm deformation and results in male infertility.General significance
Ssp411PB/PB mouse strain is an animal model of idiopathic oligoasthenoteratozoospermia (iOAT), and the gene may represent a therapeutic target for iOAT patients. 相似文献124.
Mingjun Wang Jian Wu Erye Zhou Xin Chang Jianhe Gan Tao Cheng 《Journal of cellular physiology》2019,234(11):20139-20148
125.
Ru-Jia Yan Hai-Qing Gong Pu-Ming Zhang Shi-Gang He Pei-Ji Liang 《Cognitive neurodynamics》2016,10(3):211-223
Dual-peak responses of retinal ganglion cells (RGCs) are observed in various species, previous researches suggested that both response peaks were involved in retinal information coding. In the present study, we investigated the temporal properties of the dual-peak responses recorded in mouse RGCs elicited by spatially homogeneous light flashes and the effect of the inhibitory inputs mediated by GABAergic and/or glycinergic pathways. We found that the two peaks in the dual-peak responses exhibited distinct temporal dynamics, similar to that of short-latency and long-latency single-peak responses respectively. Pharmacological studies demonstrated that the application of exogenous GABA or glycine greatly suppressed or even eliminated the second peak of the cells’ firing activities, while little change was induced in the first peak. Co-application of glycine and GABA led to complete elimination of the second peak. Moreover, application of picrotoxin or strychnine induced dual-peak responses in some cells with transient responses by unmasking a second response phase. These results suggest that both GABAergic and glycinergic pathways are involved in the dual-peak responses of the mouse RGCs, and the two response peaks may arise from distinct pathways that would converge on the ganglion cells. 相似文献
126.
克雷伯氏菌甘油脱氢酶dhaD的克隆表达、纯化及酶学性质研究 总被引:1,自引:0,他引:1
以克雷伯氏菌基因组DNA为模板,扩增得到编码甘油脱氢酶(GDH)的基因dhaD,将其克隆到大肠杆菌表达载体pET-28a(+)上,在E.coliBL21(DE3)中诱导表达,利用表达载体pET-28a(+)上的6·His-Tag标记选用Ni柱亲和层析法纯化表达具有活性的甘油脱氢酶(GDH),纯化后比酶活达到156U/mg,纯化倍数达4.6倍,回收率为67.4%。并初步研究了该酶的酶学性质,酶反应的最适pH为11.0,在pH7.0~12.0范围内稳定;酶反应的最适温度为30℃,稳定范围为25~45℃; 酶动力学参数以甘油为底物的Km为0.54 mmol/L, Vmax为0.49 μmol/(mL·min)。 相似文献
127.
陕西紫阳地区在古地理上位于扬子台地西北缘,区域内志留纪地层发育,其中志留系兰多维列统(Llandovery)特列奇阶(Telychian)笔石相地层出露较为完整,笔石带较为连续。Oktavites excentricus(Bjerreskov,1975)是特列奇阶Oktavites spiralis笔石带重要的伴生分子,在世界范围内广泛分布。文中通过对紫阳地区一系列特列奇阶剖面研究后认为,Oktavites excentricus较短的化石延限和广泛的古地理分布使其具备地层对比潜力,这将为Oktavites spiralis笔石带的进一步细分提供依据。同时,Oktavites excentricus胞管为典型的奥氏笔石式,其笔石体的盘旋方式又与稍晚的Cyrtograptus lapworthi(Tullberg,1883)(笔石带化石,弓笔石类的早期代表)十分接近,因此,对该种的演化过程的研究也为讨论弓笔石的起源过程提供了更多参考。 相似文献
128.
Liu Siqi Wang Bo Li Xiaojing Pan Jingxian Qian Xuexue Yu Yahui Xu Ping Zhu Jian Xu Xiaofeng 《Plant Cell, Tissue and Organ Culture》2019,137(3):485-494
Plant Cell, Tissue and Organ Culture (PCTOC) - Callus is a remarkable regeneration tissue. The genes correlated with root development can be involved in regulating callus development in higher... 相似文献
129.
Universal infantile hepatitis B virus (HBV) vaccination may lead to an increase in vaccine escape variants, which may pose a threat to the long-term success of massive vaccination. To determine the prevalence of occult infections in Korean vaccinated individuals, 87 vaccinated subjects were screened for the presence of HBV DNA using both the nested PCR protocol and the VERSANT HBV DNA 3.0 assay. The mutation patterns of variants were analyzed in full-length HBV genome sequences. Their HBsAg secretion and replication capacities were investigated using both in vitro transient transfection and in vivo hydrodynamic injection. The presence of HBV DNA was confirmed in 6 subjects (6.9%). All six variants had a common mutation type (X8Del) composed of an 8-bp deletion in the C-terminal region of the HBV X gene (HBxAg). Our in vitro and in vivo analyses using the full-length HBV genome indicated that the X8Del HBxAg variant reduced the secretion of HBsAg and HBV virions compared to the wild type. In conclusion, our data suggest that a novel mutation (X8Del) may contribute to occult HBV infection in Korean vaccinated individuals via a reduced secretion of HBsAg and virions, possibly by compromising HBxAg’s transacting capacity. 相似文献
130.
ABSTRACT: OBJECTIVE: To study the clinico-pathological characteristics of Langerhans cell sarcoma (LCS) which involving epidermis. METHODS: A case of primary multifocal LCS was analyzed in histopathology and immunophenotype. RESULTS: A 41-year-old man with multifocal cutaneous LCS involving the inguina and waist was reported. Clinical and pathology data were available. Neoplastic cells with markedly malignant cytological features were observed. Tumor cells exhibited irregular shape with abundant and eosinophilic red staining cytoplasm; large, irregular-shaped, showing lobulated or dented nucleus and some cells with a longitudinal nuclear groove and prominent nucleoli. The tumor cells expressed CD1a, Langerin (CD207), S-100 protein, CD68 and vimentin, and did not express pan-T or B cell markers and epithelial markers. The patient died less than 1 year after diagnosis due to local recurrence and metastasis to the lung, despite the administration of local radiation and chemotherapy. CONCLUSIONS: LCS is a tumor with markedly malignant cytological features that originates from Langerhans cells. Primary multifocal neoplasms involving epidermis is even rare. Accurate diagnosis is based on the histopathological and immunohistochemical of the tumor cells.Virtual slideThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1182345104754765. 相似文献