利用QuantStudioTM 3D数字PCR分析转基因玉米MON863含量   总被引：1，自引：0，他引：1  

胡佳莹  姜羽  杨立桃 《农业生物技术学报》2016,(8):1216-1224

QuantStudioTM 3D数字PCR (QuantStudioTM 3D digital PCR,3D-dPCR)是一种基于超高密度亲疏水微孔芯片实现数字PCR分液原理的新型核酸绝对定量平台,在转基因生物定量领域具有极大的应用前景.本研究基于3D-dPCR平台,以转基因玉米(Zea mays)MON863混合样品为例,建立基于单重和双重数字PCR体系的转基因生物(genetically modified organisms,GMOs)含量分析方法.与传统qRT-PCR比较发现,在缺乏样品纯度、纯合度信息的情况下,数字PCR能够较好地排除这些因素的影响,测定准确的量值.研究结果表明,QuantStudioTM 3D数字PCR是一种适用于转基因生物含量分析的精确定量方法,还可反映转基因玉米种子的基因型.本研究基于3D-dPCR建立的转基因玉米MON863单重和双重定量方法为转基因检测提供了新的方法和参考.  相似文献   

Whole-genome sequence of a novel Chinese cyprinid herpesvirus 3 isolate reveals the existence of a distinct European genotype in East Asia     

《Veterinary microbiology》2015,175(2-4):185-194

Cyprinid herpesvirus 3 (CyHV3), also known as koi herpesvirus (KHV), can be subdivided primarily into European and Asian genotypes, which are represented by CyHV3-U or CyHV3-I and CyHV3-J, respectively. In this study, the whole genome sequence of a novel Chinese CyHV3 isolate (GZ11) was determined and annotated. CyHV3-GZ11 genome was found to contain 295,119 nucleotides with 52.9% G/C content, which is highly similar to those of published CyHV3-U, CyHV3-I, and CyHV3-J strains. With reference to CyHV3-U, CyHV3-I, and CyHV3-J, CyHV3-GZ11 was also classified into 164 open reading frames (ORF), which include eight repeated ORFs. On the basis of the 12 alloherpeviruses core genes, results from phylogenetic analysis showed that CyHV3-GZ11 had closer evolutionary relationships with CyHV3-U and CyHV3-I than with CyHV3/KHV-J, which were also supported by genome wide-based single nucleotide substitution analysis and the use of a series of developed molecular markers. This study was the first to reveal the presence of a distinct European CyHV3 genotype in East and Southeast Asia at a whole genome level, which will evoke new insights on exploring the origin, evolution, and epidemiology of the virus.  相似文献   

Wx^mp基因背景下可溶性淀粉合成酶基因SSⅡa和去分支酶基因PUL对水稻蒸煮食味品质的影响     

姚姝  张亚东  刘燕清  赵春芳  周丽慧  陈涛  赵庆勇  朱镇  Balakrishna PILLAY  王才林 《中国水稻科学》2020,34(3):217-227

【目的】分析在同一主效基因(Wx^mp)背景下可溶性淀粉合成酶基因SSⅡa和去分支酶基因PUL对稻米蒸煮食味品质的影响,以期为水稻品质遗传改良提供依据。【方法】选择在SSⅡa和PUL存在多态性而其他淀粉合成酶相关基因没有多态性的半糯品系宁0145和粳稻品种武运粳21进行杂交,获得F2群体与F3株系。利用分子标记,选择含有Wx^mp基因的F2单株与F3株系,将这些F2单株与F3株系分成SSⅡa^nPUL^n、SSⅡa^nPUL^w、SSⅡa^wPUL^n和SSⅡa^wPUL^w4种基因型(n和w分别表示该基因来源于宁0145和武运粳21),分析不同基因型蒸煮食味品质性状的差异,探讨同一Wxmp基因背景下不同SSⅡa和PUL等位基因对蒸煮食味品质性状的影响。【结果】不同基因型间蒸煮食味品质性状均存在显著差异,来源于武运粳21的SSⅡa^w基因和PUL^w基因分别使直链淀粉含量增加0.29%~1.00%和0.62%~1.18%,且PUL的效应大于SSⅡa,两者间存在互作效应。SSⅡa^w基因和PUL^w基因降低胶稠度和崩解值,提高了热浆黏度、冷胶黏度、消减值和回复值,对糊化温度、峰值黏度和峰值时间的作用较小。【结论】明确了Wx^mp背景下SSⅡa和PUL基因对稻米蒸煮食味品质的遗传效应,该研究结果为SSⅡa和PUL基因的分子标记辅助选择改良稻米品质提供了理论依据。  相似文献   

延边黄牛ATF3基因组织表达及其多态性与肉质性状的相关性分析     

邵静  尹宝珍  焦石  张珈溯  张洛萌  耿春银  夏广军 《中国畜牧兽医》2020,47(2):488-496

  相似文献   

miR-106b-5p靶向KLF4调控山羊肌内前体脂肪细胞分化   总被引：1，自引：1，他引：0  

杜宇  赵越  林亚秋  朱江江  王永  马洁琼  谢光杰 《畜牧兽医学报》2020,51(6):1219-1228

旨在明确miR-106b-5p对山羊肌内前体脂肪细胞分化的影响，并确定这种作用是通过靶向KLF4来实现的。本研究利用实时荧光定量PCR （quantitative real-time PCR，qRT-PCR）技术检测miR-106b-5p在山羊肌内前体脂肪细胞分化过程中的表达模式，通过脂质体转染技术将miR-106b-5p mimic和miR-106b-5p inhibitor转入体外培养的山羊肌内前体脂肪细胞，油红O染色法从形态学验证miR-106b-5p对脂肪细胞中脂滴积聚的影响，qRT-PCR检测预测的靶标基因KLF4和脂肪分化标志基因的表达情况，利用双荧光素酶报告系统鉴定miR-106b-5p与KLF4的靶标关系。qRT-PCR结果显示，miR-106b-5p在山羊肌内前体脂肪细胞诱导分化第3天时表达量最高。在山羊肌内脂肪细胞中干扰miR-106b-5p后油红O染色显示脂滴聚积减少，过表达miR-106b-5p后脂滴聚积增加。在山羊肌内前体脂肪细胞中转染miR-106b-5p inhibitor后PPARγ表达量显著降低（P<0.05），而KLF4表达量极显著升高（P<0.01）；转染miR-106b-5p mimic后LPL和PPARγ表达量极显著升高（P<0.01）。荧光素酶活性试验结果显示，过表达miR-106b-5p可显著抑制KLF4荧光活性。miR-106b-5p通过靶向并负调节KLF4的表达促进山羊肌内脂肪细胞分化。  相似文献   

Sublytic C5b-9 induces protective autophagy in cultured podocytes     

LV Qian-ying  ZHOU Jian-hua  YANG Feng-jie  PU Jin-yun  ZHANG Yu 《园艺学报》2015,31(1):59-63

AIM: In podocytes, autophagy occurs at a high basal level and dysregulated autophagy is associa-ted with a variety of podocytopathies. This paper is to investigate the role of autophagy in sublytic C5b-9-induced podocyte injury. METHODS: Sublytic complement C5b-9 stimulation was used as an in vitro model. Autophagosomes were confirmed using monodansylcadaverine (MDC) staining. Immunoblotting was used to measure the change of autophagy-related markers. Cellular morphological changes were observed by Wright-Giemsa staining. Immunofluorescence staining and confocal microscopy were used to detect the expression and distribution of nephrin. The cell viability was assessed by methylthiazol tetrazolium (MTT) assay. The cell apoptosis was assessed by Annexin V-fluorescein isothiocyanate/PI staining. RESULTS: For ensuring sublytic complement injury, the maximal amounts of anti-podocyte antiserum and 160×-diluted normal human serum were used without inducing cell lysis (defined as >5% LDH release). Sublytic C5b-9 promoted autophagy of podocytes in vitro. The proautophagic effect of sublytic C5b-9 manifested in the form of accumulated MDC-labeled vesicles and enhanced the expression of LC3-Ⅱ. Autophagy inhibitor 3-methyladenosine (3-MA) promoted sublytic C5b-9-induced podocyte morphological abnormalities. Compared with the sublytic C5b-9-injured podocytes, 3-MA exposure further decreased the expression of nephrin. 3-MA enhanced sublytic C5b-9-induced podocyte apoptosis. CONCLUSION: Sublytic C5b-9 attack induces autophagy, which may play a protective role against complement-mediated podocyte injury.  相似文献   

Inlfuence of FSH Treatment on Expression of CDC25A,TSSK3 and P53in Vitro Cultured Sertoli Cells of Calf     

Li Yu-long  Wu Qiong  Zhao Xun-wu  Zeng Yue  Elkanah Adegoke  Zhang Gui-xue 《东北农业大学学报(英文版)》2015,(1)

CDC25A, TSSK3 and P53 expressionsin vitro in cultured sertoli cells after FSH treatment were studied in order to provide some data for further researches of spermatogenesis. Different concentrations of...  相似文献   

Fusarium solani Upregulated Sesquiterpene Synthase Expression,Sesquiterpene Production and Allelopathic Activity in Piper betle L.     

Kamrai Woranoot  Rattikarn Buaruang  Kodchakorn Aranyakanon  Kumrop Ratanasut  Anupan Kongbangkerd  Panatda Jannoey  Pranee Nangngam  Chonnanit Choopayak 《水稻科学》2019,26(5):290-299

Activators of sesquiterpene synthase (STS) gene expression and sesquiterpene production in Piper betle L. were examined using quantitative real time PCR and gas chromatography mass spectrometry methods, and the allelopathic activity of untreated and Fusarium solani-treated betel extracts was tested on seed germination and on the shoot and root growth of Thai rice variety PSL2 (Oryza sativa cv. Phitsanulok 2) and three dominant paddy weeds (Eclipta prostrata, Echinochloa crus-galli and Chloris barbata). The results demonstrated that F. solani dramatically upregulated STS expression and productions of β-cubebene, β-caryophyllene and germacrene D sesquiterpene when compared with the untreated control, and that betel extracts had a greater inhibitory effect on weeds than on rice. The effects were more clearly detected on seed germination and root growth than on shoot growth, and they were found to be dose-dependent. It is also noted that F. solani-treated extract had stronger effects than the untreated extract. The species most sensitive to the allelopathic effects was C. barbata, germination of which was completely inhibited even at a dose of 0.1 mg/mL untreated extract. With regards to rice, although betel extract at 1.0 mg/mL showed no inhibition on germination, it affected the elongation of rice roots, in addition to those of the tested weeds. The obtained data suggested that F. solani has potential as an activator of sesquiterpene allelochemical production via STS expression, the latter leading to the treated betel extract having a stronger phytotoxic effect. These results were beneficial in the promotion of natural herbicide production using biotechnology.  相似文献   

水产调味品鲫鱼汁的研制     

郝教敏  姚玉莲 《山西农业大学学报(自然科学版)》2011,31(6):545-550

随着人们生活水平的提高,对食品质量的要求也在不断的提高,调味品在食品质量中扮演着非常重要的角色。本实验以普通养殖鲫鱼为主要原料,研究水产调味品鲫鱼汁的最佳工艺。通过正交试验和感官评价,最终得到水产调味鲫鱼汁的最佳配方:黄原胶0.04%,鲫鱼汁30%,盐15%,糖5%,谷氨酸钠5%,5′-肌苷酸0.25%,叶黄素0.03%,鱼味香精0.03%,山梨酸钾0.05%,均质压力35MPa。水产调味品鲫鱼汁质地均匀,口感细腻,鱼鲜风味突出,色泽淡黄,气味芬芳,符合人们对于风味和营养的需求,具有广阔的市场前景。  相似文献   

HMGCR基因多态性与肉牛屠宰性状相关性分析          下载免费PDF全文

姚玉妮  宋恩亮  刘晓牧  成海建  游伟  韩红  刘桂芬 《家畜生态学报》2015,36(3)

MHGCR是胆固醇生物合成过程中的限速酶,在内源胆固醇生物合成过程中起着重要的作用。研究以牛HMGCR基因为候选基因,采用PCR-SSCP和测序技术在利杂牛、利西杂牛、秦川牛、西杂牛和夏西杂牛五个群体共123个个体中检测遗传变异情况。结果表明,检测到AA和AB共2种基因型,没有发现BB基因型,测序分析发现一处SNP多态位点(GenBank:BC153262.1 11137TC)。不同基因型与屠宰性状的相关性分析显示,在利西杂群体中AA基因型个体与AB基因型个体相比,AA基因型个体的屠宰率和牛柳指标达到了极显著水平(P0.01),而AA基因型的上脑重显著高于AB基因型(P0.05),其余各项指标基因型间差异不显著(P0.05)。因此,HMGCR基因单核苷酸多态性对利西杂牛脂肪沉积有一定的影响,可作为影响屠宰性状的候选基因,指导肉牛的育种工作。  相似文献