排序方式: 共有11条查询结果,搜索用时 15 毫秒
1.
传统辣椒DNA提取常采用CTAB法,该技术已难满足大量开展分子生物学工作的需要,为了解决此困境,进行了辣椒上碱煮法提取的尝试。将待测物置PCR管中,向管中加入80μL 0.2 mol/L Na OH溶液,置PCR仪上以100℃处理30 min,再以40μL Tris-HCl缓冲液将PCR管中溶液调至p H 7~8后,以之为模板可在后续的SSR-PCR中得到良好的实验结果。结果表明针对不同的研究目的需采取不同的组织部位来进行DNA的提取,当面临种子纯度测定的任务时,需要截取种子的芽根。当面临成熟植株的基因型筛选的任务时,需要截取植株的顶端嫩叶。综上,本研究表明相较其他传统的DNA提取方法,碱煮法确是更适合解决辣椒大规模鉴定问题的技术手段。 相似文献
2.
庞泉沟国家级自然保护区森林景观格局动态 总被引:3,自引:1,他引:2
以1990年TM,1999年ETM和2004年SPOT5等3期卫星影像数据为基础资料,应用遥感技术(RS)和地理信息系统(GIS)技术,通过斑块密度、多样性指数、均匀度指数和聚合度等多种指标,对山西省庞泉沟国家级自然保护区的森林景观格局动态进行全面分析。结果表明:研究时段内的景观总体的斑块密度和边缘密度持续增加,表明景观总体的破碎化程度增加。景观整体的多样性有所下降,均匀度指数也处于低水平,说明景观中的优势类型逐渐加强其优势地位。保护区管理部门应密切关注景观破碎化现象,建议调整经营措施,控制人为干扰,延长采伐周期,增加林分郁闭度。表4参12 相似文献
3.
The total DNA was extracted from strawberry leaves infected with Strawberry vein banding virus (SVBV) by CTAB method. Specific primer pairs were designed to amplify the gene ORF I of SVBV-Shenyang isolate by PCR, gene ORF I was cloned into modified prokaryotic expression vector pET-32a (+)-GST, the recombinant plasmid pET-ORF I was transformed into E. coli DH5α, then the positive clones were screened and sequenced. The recombinant plasmid was extracted and transformed into Escherichia coli BL2l (DE3), the fusion protein with an approximate molecular weight of 56 kDa was obtained with IPTG induction and Ni2+-NTA affinity column purification. The purified fusion protein was used to immunize the rabbits to prepare the specific antiserum. The result of ELISA showed that the titer of the prepared antiserum is up to 1:520 000, Western blot analysis indicated that the prepared antiserum could reacted specifically with purified recombinant fusion protein. Not only the expression of P1 protein in SVBV infected-strawberry leaves, but also the expression of P1 protein in P1-infiltrated Nicotiana benthamiana leaves could be detected, using 2 000 times diluted antiserum. 相似文献
4.
为了准确快速建立薄皮甜瓜品种翡翠的SSR鉴定体系,利用234对甜瓜SSR特异性引物对翡翠品种亲本进行引物筛选,筛选出3对扩增出差异片段的引物,结合商品种F1代进行验证,获得1条最优引物.在开展田间人为掺杂母本并定植后,大田和室内2种鉴定方法对同一定植苗开展一一对应鉴定试验,结果发现室内分子鉴定与大田鉴定符合率达100%;再对生产销售近2 a的样本鉴定发现,室内鉴定与大田鉴定符合率达99%以上.结果表明,薄皮甜瓜品种翡翠的SSR快速鉴定技术可以应用于实际生产中. 相似文献
5.
传统的辣椒DNA提取常采用CTAB法,该技术已难满足大量开展分子生物学工作的需要,为了解决此困境,进行了辣椒上碱煮法提取的尝试。一个可用于新品种的SSR分子标记被用作检验手段,验证碱煮法所提DNA是否可满足实验需要。结果表明针对不同的研究目的需采取不同的组织部位来进行DNA的提取,当面临种子纯度测定的任务时,需要截取种子的芽根。当面临成熟植株的基因型筛选的任务时,需要截取植株的顶端嫩叶。以上这些植物材料置PCR管中,在向管中加入80 μL 0.2 mol/L NaOH溶液后,置PCR仪上以100℃处理30分钟,
再以40 μL Tris-HCl缓冲液将pH值调节至7~8之间后,以之为模板可在后续的SSR-PCR中得到良好的实验结果。综上,本研究表明相较其他耗时耗力的传统DNA提取方法,碱煮法确是更适合解决辣椒大规模鉴定问题的技术手段。 相似文献
6.
The total DNA was extracted from strawberry leaves infected with Strawberry vein banding virus (SVBV) by CTAB method. Specific primer pairs were designed to amplify the gene ORF I of SVBV-Shenyang isolate by PCR, gene ORF I was cloned into modified prokaryotic expression vector pET-32a (+)-GST, the recombinant plasmid pET-ORF I was transformed into E. coli DH5α, then the positive clones were screened and sequenced. The recombinant plasmid was extracted and transformed into Escherichia coli BL2l (DE3), the fusion protein with an approximate molecular weight of 56 kDa was obtained with IPTG induction and Ni2+-NTA affinity column purification. The purified fusion protein was used to immunize the rabbits to prepare the specific antiserum. The result of ELISA showed that the titer of the prepared antiserum is up to 1:520 000, Western blot analysis indicated that the prepared antiserum could reacted specifically with purified recombinant fusion protein. Not only the expression of P1 protein in SVBV infected-strawberry leaves, but also the expression of P1 protein in P1-infiltrated Nicotiana benthamiana leaves could be detected, using 2 000 times diluted antiserum. 相似文献
7.
8.
9.
10.
【目的】探讨南方水稻黑条矮缩病毒(Southern rice black-streaked dwarf virus,SRBSDV)安徽分离物(SRBSDVAn Hui-HN2)的遗传特性,并获得原核表达的P5-2蛋白。【方法】RT-PCR扩增SRBSDV S5片段,克隆、测序并进行序列分析。将S5-2基因插入原核表达载体,重组载体转化大肠埃希菌并用IPTG诱导,Ni2+-NTA亲和柱纯化融合蛋白,SDS-PAGE分析P5-2蛋白的表达情况。【结果】SRBSDV-An Hui-HN2 S5片段全长3 167 bp,包含S5-2基因全长612 bp,编码204个氨基酸。序列比对结果显示,SRBSDV-An Hui-HN2 S5片段与其他SRBSDV分离物S5片段的序列相似性极高,达99.0%~99.7%,而与斐济病毒属(Fijivirus)其他成员S5片段的序列相似性较低,仅为38.0%~71.3%;构建的S5片段系统发育树表明SRBSDV和RBSDV聚成1个分支,其中6个SRBSDV分离物聚成1个亚分支。原核表达获得相对分子质量约为47 000的重组蛋白,Western blot分析显示,GST单抗能够与重组融合蛋白发生特异性反应。【结论】SRBSDV各分离物之间亲缘关系非常近,而与Fijivirus其他成员亲缘关系较远,原核表达获得的融合蛋白为靶标蛋白。 相似文献