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苹果细菌人工染色体双色DNA纤维荧光原位杂交体系的建立及应用 被引量:1
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作者 王三红 章镇 +1 位作者 蔡斌华 渠慎春 《中国农业科学》 CAS CSCD 北大核心 2014年第11期2205-2213,共9页
【目的】建立苹果DNA纤维荧光原位杂交(Fiber FISH)技术体系,从而为利用该技术确定苹果基因组DNA序列间的位置关系、构建精细物理图谱等方面的应用奠定基础。【方法】以‘Florina’苹果幼叶为试材,通过液氮研磨,尼龙膜过滤和TritonX-10... 【目的】建立苹果DNA纤维荧光原位杂交(Fiber FISH)技术体系,从而为利用该技术确定苹果基因组DNA序列间的位置关系、构建精细物理图谱等方面的应用奠定基础。【方法】以‘Florina’苹果幼叶为试材,通过液氮研磨,尼龙膜过滤和TritonX-100去除叶绿素等步骤提取细胞核。细胞核经碱裂解,采用盖玻片拉伸方法制备DNA纤维。比较细胞核不同裂解时间和在5种不同包被类型的载玻片上DNA纤维拉伸的效果。用碱裂解方法提取来自苹果‘Florina’自交不亲和S9基因座的4个细菌人工染色体(Bacterium Artificial Chromosome)BAC 34G16、BAC 45M19、BAC 70J19和BAC 69A4。提取的质粒经PEG纯化,用地高辛或生物素标记探针。探针与DNA纤维制片经过80℃变性、37℃杂交2—3 d和洗片,采用"三明治"方法进行信号放大和检测,在荧光显微镜下观察试验结果。【结果】建立了以苹果幼叶为材料,液氮研磨提取细胞核,碱裂解细胞核和盖玻片拉伸制备DNA纤维的试验方法。提取的细胞核纯净、结构完整,细胞核浓度>5×103个/μL。试验结果表明,细胞核裂解4 min,在多聚赖氨酸包被的载玻片上制备的DNA纤维平直、伸展均匀,纤维量多、细长,效果好。经原位杂交和信号检测,获得了清晰的、具有Fiber FISH典型特征的"念珠状"杂交信号。对已知大小和位置关系的两个BAC克隆BAC 34G16和BAC 45M19进行杂交信号测量和分析,得出BAC克隆大小(Y,Kb)与信号长度(X,μm)的相关方程为Y=3.47X(R2=0.9215),其斜率即为该试验技术体系Fiber FISH的分辨率3.47 kb·μm-1。试验对未知大小和位置关系的两个BAC克隆BAC 70J19和BAC 69A4成功地进行了鉴定,它们大小分别为(112.1±18.4)kb和(133.2±16.3)kb,之间有(90.2±7.3)kb的重叠区域。【结论】建立了以苹果幼叶提取细胞核,制备DNA纤维和原位杂交的方法,获得了高分辨率的苹果DNA纤维原位杂交试验技术体系。 展开更多
关键词 苹果 细菌人工染色体 DNA纤维荧光原位杂交
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大麦45S和5SrDNA定位及5SrDNA伸展纤维的FISH分析 被引量:14
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作者 赵丽娟 李立家 +2 位作者 覃瑞 熊怀阳 宋运淳 《武汉植物学研究》 CSCD 北大核心 2005年第1期15-19,共5页
用荧光原位杂交技术对45S和5SrDNA在大麦(HordeumvulgareL.)有丝分裂中期染色体进行了确定分析,较强的45SrDNA信号共有2对,分别分布在大麦的第1染色体的短臂和第2染色体的长臂。而5SrDNA则只有1对杂交信号,位于第3染色体的长臂,但信号... 用荧光原位杂交技术对45S和5SrDNA在大麦(HordeumvulgareL.)有丝分裂中期染色体进行了确定分析,较强的45SrDNA信号共有2对,分别分布在大麦的第1染色体的短臂和第2染色体的长臂。而5SrDNA则只有1对杂交信号,位于第3染色体的长臂,但信号较弱。用伸展DNA纤维的荧光原位杂交(Fiber-FISH)技术测定了5SrDNA在大麦的基因组中的拷贝数,计算出5SrDNA的拷贝数约为408~416。对大麦品种中rDNA位点数目的可变性进行了讨论。 展开更多
关键词 大麦 RDNA FISH定位 DNA纤维荧光原位杂交
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玉米DNA纤维的制备及端粒DNA的Fiber-FISH
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作者 赵丽娟 刘良科 李立家 《安徽农业科学》 CAS 北大核心 2008年第6期2251-2252,2258,共3页
[目的]建立了一种快速制备DNA纤维的方法,用端粒DNA在玉米纤维上进行物理定位。[方法]采用刀切法从玉米嫩叶中提取细胞核。以端粒DNA为探针,在玉米DNA纤维上进行伸展DNA纤维的荧光原位杂交(Fiber-FISH),研究端粒DNA重复序列在玉米染... [目的]建立了一种快速制备DNA纤维的方法,用端粒DNA在玉米纤维上进行物理定位。[方法]采用刀切法从玉米嫩叶中提取细胞核。以端粒DNA为探针,在玉米DNA纤维上进行伸展DNA纤维的荧光原位杂交(Fiber-FISH),研究端粒DNA重复序列在玉米染色体上的拷贝数。[结果]用刀切法提取玉米细胞核,提高了核的完整性,并改善了DNA纤维的制备效果。玉米细胞核裂解的最佳时间为8-9 min。玉米的Fiber-FISH试验结果表明,杂交信号为伸展的念珠状长链,玉米各条染色体端粒DNA的长度为7-103μm,各染色体端粒重复序列的拷贝数存在显著差异(为15~230 kb)。[结论]玉米各染色体中端粒的长度可能不同,而且随着玉米生长及环境的变化,各条染色体端粒DNA长度的变化也不一致。 展开更多
关键词 玉米 端粒DNA纤维荧光原位杂交 拷贝数
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Breeding of Transgenic Hybrid Cotton Variety‘Yumian 50' and Cultivation Technology
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作者 唐中杰 房卫平 +2 位作者 吕淑平 聂利红 谢德意 《Agricultural Science & Technology》 CAS 2017年第8期1399-1402,共4页
The mid-early-maturing new cotton variety 'Yumian 50' was bred by In-stitute of Industrial Crops, Henan Academy of Agricultural Sciences. It was exam-ined and approved by Henan Provincial Crop Variety Approval Commi... The mid-early-maturing new cotton variety 'Yumian 50' was bred by In-stitute of Industrial Crops, Henan Academy of Agricultural Sciences. It was exam-ined and approved by Henan Provincial Crop Variety Approval Committee in 2016 (Yushenmian 2016002). In order to study the variety characteristics of 'Yumian 50' and the key points of its cultivation technology and accelerate its application in cot-ton production, the breeding process of 'Yumian 50' was described in this paper. The test data of 26 times for 3 years of 'Yumian 50' participating in Henan region-al tests in 2012-2014 were summarized. The results showed that 'Yumian 50' had outstanding performance in Henan regional tests and production tests. 'Yumian 50' is a new cotton variety with characteristics of high yield, good quality and high re-sistance to diseases and pests. 展开更多
关键词 Transgenic Hybrid Cotton 'Yumian 50' YIELD Fiber Quality CultivationTechnoloav
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Tensile properties of strain-hardening cementitious composites containing polyvinyl-alcohol fibers hybridized with polypropylene fibers 被引量:4
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作者 H.R.Pakravan M.Jamshidi 《Journal of Central South University》 SCIE EI CAS CSCD 2018年第1期51-59,共9页
Partially replacing polyvinyl-alcohol(PVA)fibers with polypropylene(PP)fibers in strain-hardening cementitious composites(fiber hybridization)modify certain mechanical properties of these materials.The hybridization b... Partially replacing polyvinyl-alcohol(PVA)fibers with polypropylene(PP)fibers in strain-hardening cementitious composites(fiber hybridization)modify certain mechanical properties of these materials.The hybridization based on the introduction of low-modulus hydrophobic polypropylene fibers improves the ductility and the strain-hardening behavior of the cementitious composites containing polyvinyl-alcohol fibers of different types(PVA-SHCC).Pull-out tests indicate that adding PP fibers increases the energy capacity of the hybrid composite with respect to the material containing only PVA fibers under tensile loading,and PP-fiber geometry(i.e.,section shape and length)is a key factor in enhancing the strain capacity. 展开更多
关键词 polyvinyl-alcohol (PVA) strain-hardening cementitious composite (PVA-SHCC) fiber hybridization
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Preparation and Detection of Extended DNA Fibers of Chinese Cabbage
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作者 李九欢 王彦华 +2 位作者 李晓峰 申书兴 轩淑欣 《Agricultural Science & Technology》 CAS 2012年第3期517-519,591,共4页
Abstract [Objective] The paper was to prepare and detect the extended DNA fibers of Chinese cabbage. [Method] Chinese cabbage nuclei was first successfully isolated by chopping young leaves with a blade, then nuclei w... Abstract [Objective] The paper was to prepare and detect the extended DNA fibers of Chinese cabbage. [Method] Chinese cabbage nuclei was first successfully isolated by chopping young leaves with a blade, then nuclei were lysed by SDS to release DNA, and DNA fibers were dragged and extended with a coverslip. [Result] The results of Fiber-FISH with genomic DNA and 25S rDNA as probes showed that DNA fiber size as long as about 1.93 Mb could be measured and the number of 25S rDNA copies region were estimated to be 258 and 687 in Chinese cabbage genome. DNA fibers prepared by this method showed equally spread parallel thread with clear background, and were suitable for FISH analysis. [Conclusion] The study would accelerate Chinese cabbage genome mapping and organization analysis. 展开更多
关键词 Chinese cabbage Nuclei isolation Extended DNA fibers (EDFs) Fluorescence in situ hybridization (FISH) 25S rDNA
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Preliminary evaluation of five elephant grass cultivars harvested at different time for sugar production 被引量:1
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作者 李媛媛 张叶龙 +4 位作者 郑洪波 杜健 张红漫 吴娟子 黄和 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2015年第7期1188-1193,共6页
Five elephant grass cultivars, Pennisetum purpureum, cv. Huanan (Huanan), P. purpureum, cv. N51 (N51), P. purpureum, cv. Sumu No. 2 (Sumu-2), ( Penniseturn americanum x P. purpureum ) x P. purpureum cv. Guimu ... Five elephant grass cultivars, Pennisetum purpureum, cv. Huanan (Huanan), P. purpureum, cv. N51 (N51), P. purpureum, cv. Sumu No. 2 (Sumu-2), ( Penniseturn americanum x P. purpureum ) x P. purpureum cv. Guimu No. 1 (Guimu-1) and P. americanum cv. Tift23A x P. purpureum cv. Tilt NS1 (Hybrid Pennisetum), at three harvest stages were studied. With dilute sulfuric acid pretreatment followed by enzymatic hydrolysis, it is found that cel- lulose conversion of the five elephant grass cultivars harvested in August and September is higher than that har- vested in October. The cellulose conversion for elephant grass cultivars harvested in August and September follows an order of Hybrid Pennisetum 〉 Sumu-2 〉 Huanan 〉 Guimu-1 〉 N51. This may be explained by the fact that lignification is gradually strengthened with time, inhibiting degradation of cellulose and hemicellulose. Moreover, cellulose conversions of Hybrid Pennisetum, Sumu-2 and Huanan harvested in August and September are higher based on hierarchical clustering results. 展开更多
关键词 Energy crop Elephant grass Dilute sulfuric acid pretreatment Enzymatic hydrolysis Glucose
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水稻5S rDNA和着丝粒顺序RCS2拷贝数的Fiber-FISH测定 被引量:2
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作者 李宗芸 黄思罗 +3 位作者 金危危 宁顺斌 宋运淳 李立家 《科学通报》 EI CAS CSCD 北大核心 2001年第19期1641-1644,T004,共5页
用伸展DNA纤维的荧光原位杂交(Fiber-FISH)技术测定了5S rDNA和着丝粒DNA顺序RCS2在水稻广陆矮四号(Oryza sativa ssp indica cv Guangluai No4)基因组中的拷贝数. 为了确定拷贝数, 需要知道显微镜下一定长度DNA纤维所含碱基对数. 为此,... 用伸展DNA纤维的荧光原位杂交(Fiber-FISH)技术测定了5S rDNA和着丝粒DNA顺序RCS2在水稻广陆矮四号(Oryza sativa ssp indica cv Guangluai No4)基因组中的拷贝数. 为了确定拷贝数, 需要知道显微镜下一定长度DNA纤维所含碱基对数. 为此, 测量了两个已知碱基对数DNA序列的伸展纤维在显微镜下的长度. 其中供试BAC38D17的插入序列为136 kb, 其伸展纤维在显微镜下的长度为56.4 μm, 平均为2.41 kb/μm; BAC44B4全长为144.5 kb, 其伸展纤维的长度为55.7 μm, 平均为2.60 kb/μm. 这与Watson-Crick模型中B-DNA的2.97 kb/μm十分接近. 根据两个样本每微米平均碱基数, 即2.51 kb/μm的标准, 计算出5S rDNA的拷贝数约为686, 着丝粒DNA顺序RCS2约为286~1121拷贝. 展开更多
关键词 着丝粒DNA顺序 5S RDNA 拷贝数 DNA纤维荧光原位杂交 DNA Fiber-FISH 水稻 碱基对数
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A localized surface plasmon resonance DNA biosensor based on gold nanospheres coated on the tip of the fiber
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作者 贾朔 边超 +2 位作者 佟建华 孙楫舟 夏善红 《Optoelectronics Letters》 EI 2016年第2期157-160,共4页
A localized surface plasmon resonance(LSPR) biosensor was prepared with gold nanospheres(Au NSs) coated on the tip face of the optical silica fiber. Au NSs with the sizes of 20 nm and 80 nm were used. The sensitivitie... A localized surface plasmon resonance(LSPR) biosensor was prepared with gold nanospheres(Au NSs) coated on the tip face of the optical silica fiber. Au NSs with the sizes of 20 nm and 80 nm were used. The sensitivities of Au NS_(20 nm) and Au NS_(80 nm) modified sensors to bulk refractive index(RI) variation are 82.86 nm/RIU and 218.98 nm/RIU, respectively. The Au NS_(80 nm) modified sensor was used for the detection of 40 bases DNA hybridization and the limit of detection is 50 nmol/L, where the 40-bases DNA probe was covalently linked with Au NS_(80 nm). The complementary DNA sequence in tris-acetate-EDTA(TAE) buffer solution was detected as the target DNA. This fiber sensor has the advantages of small sample consumption, easy fabrication and high sensitivity. 展开更多
关键词 biosensor DNA coated plasmon covalently bases fabrication silica localized sizes
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