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1.
For investigating the possibility of applying degenerate oligonucleotide primer PCR (DOP-PCR) and comparative genomic hybridization
(CGH) technique to analyses of genomic genetics in a single cell, the whole genomic DNA of a single cell with XX, XY, XO,
XXY, +13 or +21 was amplified by DOP-PCR. Single cell DOP-PCR CGHs with conventional and modified control references, the
genomic DNA and a single cell DOP-PCR product from normal male, were carried out respectively. The results showed that the
average profile of the fluorescence intensity ratio in CGH with the genomic DNA as reference fluctuates much and that the
standard deviation in about 30% haploid is beyond the normal limits. False positive hyper-representation was found to exist
in X chromosome while trisomy 13 and 21 were not detected. However, the distributions of the mean and the standard deviation
of the ratio in the CGH with DOP-PCR product as reference were quite acceptable. The copy number changes of chromosome X,
Y, 13 and 21 were revealed. Those results suggested that there is unrandom unequal amplification in a single cell DOP-PCR.
Using a single DOP-PCR product as reference can decrease its influence on CGH. Single cell DOP-PCR-CGH and its application
in the genetic analyses of preimplantation embryo or fetal cell in maternal blood may be possible. 相似文献
2.
单核苷酸多态性（SNP）是指不同个体DNA序列上的单个碱基的差异,是人类基因组中最丰富的遗传变异。单体型是指位于一条染色体上或某一区域的一组相关联的SNP等位基因。研究表明在复杂性疾病研究方面,由多个变异位点组合构成的单体型所携带的信息比单个的SNP数据的信息更有价值,由此衍生了单体型装配问题。文章论述了SNP,单体型,基因型的定义,综述了求解单一个体单体型装配问题的主要模型及算法,同时阐述了求解群体单体型装配问题的5种方法及算法。 相似文献
3.
从假单胞菌（Pseudomonassp．）XZG36中克隆弹性蛋白酶基因,构建原核表达载体,实现其在大肠杆菌（Escherichiacoli）中的高效表达,并对表达产物进行酶学性质分析,为微生物发酵生产弹性蛋白酶奠定基础．以假单胞菌基因组DNA为模板,PCR扩增弹性蛋白酶基因,并将其开放阅读框（0RF）克隆至融合表达载体pET30a（＋）进一步IPTG诱导表达;表达产物经His·Bind亲和层析纯化后对弹性蛋白酶进行酶学性质分析．实验成功克隆了弹性蛋白酶基因,DNA基因片段为1672bp、编码497个氨基酸残基的多肽,与预计长度相符合;实现了其在E．coli中的高效表达,表达量约占菌体总蛋白的20％;经SDS-PAGE分析,相对分子质量为48000,与预期的一致;提纯后的表达蛋白SDS-PAGE分析可见单一条带,纯度可达92％以上．表达蛋白具有良好活性． 相似文献
4.
5.
Sun Jie Huang He Zhu Yuan-yuan Lan Jian-ping Li Jing-yuan Lai Xiao-yu Yu Jian 《Journal of Zhejiang University. Science. B》2005,6(12):1141-1147
Objective: Detecting the expression and mutation of human telomeric repeat binding factor (hTRF1) in 10 malignant hematopoietic cell line cells on the base of determining its genomic structure and its four pseudogenes
to clarify if hTRF1 mutation is one of the factors of the activation of telomerase. Methods: hTRF1cDNA sequences were obtained from GenBank, its genome structure and pseudogenes were forecasted by BLAST and other biology
information programs and then testified by sequencing. Real-time RT-PCR was used to detect the expression of hTRF1mRNA in 10 cell line cells, including myelogenous leukemia cell lines K562, HL-60, U-937, NB4, THP-1, HEL and Dami; lymphoblastic
leukemia cell lines 6T-CEM, Jurkat and Raji. Telomerase activities of cells were detected by using telomeric repeat amplification
(TRAP)-ELISA protocol. PCR and sequencing were used to detect mutation of each exon of hTRF1 in 10 cell line cells. Results: hTRF1 gene, mapped to 8q13, was divided into 10 exons and spans 38.6 kb. Four processed pseudogenes of hTRF1 located on chromosome 13, 18, 21 and X respectively, was named as ψhTRF1-13, ψhTRF1-18, ψhTRF1-21 and ψhTRF1-X respectively. All cell line cells showed positive telomerase activity. The expression of hTRF1 was significantly lower in malignant hematopoietic cell lines cells (0.0338, 0.0108:_0.0749) than in normal mononuclear cells
(0.0493, 0.0369:_0.128) (P=0.004). But no significant mutation was found in all exons of hTRF1 in 10 cell line cells. Four variants were found in part of intron 1, 2 and 8 of hTRF1. Their infection on gene function is unknown and needs further studies. Conclusion: hTRF1 mutation is probably not one of the main factors for telomerase activation in malignant hematopoietic disease.
Project supported by the National Basic Research Program (973) of China (No. 2002CB713700) and the National Natural Science
Foundation of China (No. 39870339) 相似文献
6.
几种药用植物基因组DNA提取方法研究 总被引：3，自引：0，他引：3
采用改良2×CTAB法和改良SDS法提取富含多糖和次生代谢物质的几种药用植物不同器官基因组DNA,并用紫外分光光度计分析,凝胶电泳和PCR扩增进行鉴定。结果表明:改良2×CTAB法和改良SDS法均能有效去除不同药用植物材料中的蛋白质、多糖、酚类及其他次生代谢物质,获得较高质量基因组DNA。但与改良SDS法相比,改良2×CTAB法提取的基因组DNA质量更好。 相似文献
7.
采集20例南阳牛的血液,采用酚/氯仿法分离白细胞提取基因组DNA,并且对ANGPTL4基因部分序列(677～998 bp)PCR扩增条件进行了优化。结果表明,获得的牛基因组DNA经0.8%琼脂糖凝胶电泳检测,主条带清晰,表明获得的南阳牛基因组DNA可以用于后续实验;以所提取的基因组DNA为模板,优化ANGPTL4基因片段扩增的条件,结果表明,最理想的PCR扩增条件是模板量为2μL(50 mg/L),Taq聚合酶(5 u/μL)的量为0.8μL,退火温度为56℃,循环次数为35次。 相似文献
8.
都支杜鹃ISSR扩增条件的优化 总被引：1，自引：0，他引：1
对珍稀濒危物种都支杜鹃的ISSR扩增体系中的Mg2+浓度、dNTP浓度、Taq酶含量和底物含量进行优化,并在优化后的基础上筛选出8条效果较好的引物,在梯度PCR仪中设置温度梯度,找到这些引物的最佳退火温度。优化结果表明,至少在一定的范围内底物浓度和Taq酶含量对PCR反应结果影响不大,相对而言,dNTP浓度和Mg2+浓度对反应结果的影响更加显著。优化后的15 ul PCR反应体系中包括20 ng模板DNA、0.3μM引物、1.5 ul Buffer(Mg2+free),0.5 U Taq酶、1.5 mM MgCl2和0.1 mM dNTP。 相似文献
9.
Arthur L. Beaudet 《Child development》2013,84(1):121-132
Chromosomal microarray analysis (CMA) has emerged as a powerful new tool to identify genomic abnormalities associated with a wide range of developmental disabilities including congenital malformations, cognitive impairment, and behavioral abnormalities. CMA includes array comparative genomic hybridization (CGH) and single nucleotide polymorphism (SNP) arrays, both of which are useful for detection of genomic copy number variants (CNV) such as microdeletions and microduplications. The frequency of disease‐causing CNVs is highest (20%–25%) in children with moderate to severe intellectual disability accompanied by malformations or dysmorphic features. Disease‐causing CNVs are found in 5%–10% of cases of autism, being more frequent in severe phenotypes. CMA has replaced Giemsa‐banded karyotype as the first‐tier test for genetic evaluation of children with developmental and behavioral disabilities. 相似文献
10.
根据GenBank发布的基因序列,设计PCR引物,分别从诸葛菜(Orychophragmus violaceus)的花瓣基因组DNA和cDNA克隆到查尔酮合成酶基因,并定名为OvCHS,序列已上传至NCBI数据库,登陆号为EF408918。序列分析表明,OvCHS基因的基因组全长为1263 bp,具一个75 bp的内含子,编码区全长为1188 bp,编码395个氨基酸。与模式植物拟南芥(Arabidopsis thaliana)查尔酮合成酶基因AtCHS比较发现,两基因编码区有135个碱基不同,相似性为88.64%,氨基酸序列中仅16个氨基酸残基的差异,相似性达95.95%。 相似文献
11.
ＩＮＴＲＯＤＵＣＴＩＯＮＰＦＣｃｉｒｃｕｉｔｒｙｓｉｍｕｌａｔｉｏｎｈａｓｓｏｍｅｉｎｈｅｒｅｎｔｄｉｆｆｉｃｕｌｔｉｅｓ,ａｓｔｗｏｆｒｅｑｕｅｎｃｉｅｓｈａｖｅｔｏｂｅｃｏｎ ｓｉｄｅｒｅｄｄｕｒｉｎｇｓｉｍｕｌａｔｉｏｎ :ｏｎｅｉｓｔｈｅｓｗｉｔｃｈｉｎｇｆｒｅｑｕｅｎｃｙｏｆａｂｏｕｔ 50ｋＨｚ,ａｎｏｔｈｅｒｉｓｔｈｅｍａｉｎｓｆｒｅｑｕｅｎｃｙｏｆ50ｔｏ 60Ｈｚ.Ｉｔｃａｕｓｅｓａｎｅｘｔｒｅｍｅｌｙｌａｒｇｅｄｙｎａｍｉｃｏｐｅｒａｔｉｏｎｒａｎｇｅ .Ｔｏｓｕｒｍｏｕｎｔｔｈｅｓｅｄｉｆ… 相似文献
12.
ＩＮＴＲＯＤＵＣＴＩＯＮＢｒｏａｄｂｅａｎｗｉｌｔｖｉｒｕｓ (ＢＢＷＶ)ｉｓｔｈｅｔｙｐｅｍｅｍｂｅｒｏｆｔｈｅｇｅｎｕｓＦａｂａｖｉｒｕｓ.Ｉｔｈａｓａｗｉｄｅｈｏｓｔｒａｎｇｅａｍｏｎｇｄｉｃｏｔｙｌｅｄｏｎｓａｎｄｓｏｍｅｆａｍｉｌｉｅｓｏｆｍｏｎｏｃｏｔｙｌｅｄｏｎｓ,ａｎｄｉｓａｎｅｃｏｎｏｍｉｃａｌｌｙｉｍ ｐｏｒｔａｎｔｖｉｒｕｓｉｎＣｈｉｎａ (Ｚｈｏｕｅｔａｌ.,1 994 ) .ＢＢＷＶｈａｓｉｓｏｍｅｔｒｉｃｐａｒｔｉｃｌｅｓ,ｈｅｘａｇｏｎａｌｉｎｏｕｔ ｌｉｎｅａｎｄ 3 0ｎｍｉｎｄｉａｍ… 相似文献
13.
针禾属植物羽毛针禾基因组DNA提取方法的研究 总被引：3，自引：0，他引：3
传统的CTAB DNA提取法步骤多.较烦琐,DNA产率低,而且由于酚、单宁、色素、多糖很难完全去除,容易影响随机扩增多态、微卫星等标记工作的效率.采用SDS裂解法提取了针禾属植物幼嫩叶片的基因组DNA,所获得的DNA可用于PCR反应.并且在针禾属植物的研究中得到了较好的结果. 相似文献
14.
Vani Brahmachari 《Resonance》1997,2(3):48-57
The pattern of inheritance of genes linked to the sex chromosomes in humans have their own signature due to the presence of a single copy of X and Y chromosomes in males and 2 copies of the X chromosome in females. However nature has adopted ingenious methods to equalize the copy number of most genes that map to the X chromosome. This in turn has contributed to the understanding of regulatory phenomena involving whole chromosomes. 相似文献
15.
徐忠东 《安徽教育学院学报》2007,25(6):82-85
单细胞凝胶电泳被广泛应用于遗传毒理学、DNA损伤与修复的检测、环境污染检测、细胞凋亡机制的研究等诸多领域。四膜虫是一种真核单细胞原生动物，对环境中有毒物质的毒性反应敏感，是环境污染早期预报的理想指示物种。目前已作为标准检测生物被广泛应用于毒性检测、细胞凋亡和水质检测中。本文探讨将四膜虫作为简单的模型细胞，与检测DNA损伤与修复极敏感的单细胞凝胶电泳结合起来，通过改进方法，建立一种更为灵敏、快速、简便、经济的环境污染评价方法。 相似文献
16.
探究烤鸭储存过程中菌群的变化,为延长其贮存期奠定理论基础．用酚、氯仿、异戊醇法提取烤鸭肉中细菌的基因组DNA,PCR扩增16SRNAV3区,用DGGE电泳技术分析菌群结构的变化情况,然后用Bionumerics软件对DGGE分子指纹图谱进行烤鸭肉菌群结构相似性分析．PCR结果表明,成功扩增了细菌基因组DNA,为230bp;DGGE实验结果表明,DGGE分子指纹图谱上条带存在显著差异;用bionumberics软件分析,发现样本相似度不断变化,表明在储存过程中,烤鸭肉菌群结构发生了变化．该研究为延长烤鸭肉保质期提供理论参考依据． 相似文献
17.
目的是建立一种简便、快捷的脆性X染色体综合征的临床检测方法.在常规PCR的基础上,采用热启动法,同时加入PCR增强剂Btaine、DMSO,对20例表型正常人群外周血样本进行FMR1基因（CGG）n重复序列检测.结果表明,改良后的FMR1基因PCR扩增技术能够高效扩增CG富集区,采用该方法我们检测了20例外周血样本均获得目标片段,PCR扩增产物片段长度400～500bp（相当于n=20～53）.因此,该方法简便、快速、经济而且重复性好,可用于群体普查和门诊快速筛查脆性X染色体综合征. 相似文献
18.
文章在中国当代军事文学和新时期军事文学的谱系中 ,对莫言的军事小说进行了审美观照、深度剖析和文化解码。认为莫言的军事小说在消解崇高的“非英雄化”的美学表述中出现了严重的认识论上的滑坡。因为军事文学最大的审美魅力永远在于它动人魂魄的庄严感、神圣感、使命感和崇高感 ,英雄主义、爱国主义、乐观主义是它永恒的主旋律 ,牺牲和奉献精神是它永远的坚定支点 ,任何突破与超越都必须以上述基本特征为前提。 相似文献
19.
大学与社会共生:地方高校发展的模式选择——从美国相互作用大学看我国地方高校的发展 总被引：2，自引：0，他引：2
美国相互作用大学是适应新经济时代的产物 ,其核心理念是“以他方为中心” ,与社会共谋发展。我国地方高校可借鉴其经验 ,结合本地本校的实际 ,采取独建型、依托型、改建型、校企型、远程型等模式发展自己 相似文献
20.
油松种子胚乳DNA的提取 总被引：7，自引：0，他引：7
运用改良的CTAB法,从油松萌发种子的胚乳中提取到较高质量的DNA。为从松杉类植物的种子中提取基因组DNA或从胚乳中提取单倍体DNA提供了经济、快速和可靠的实验方法,也为进一步的分子生物学分析包括PCR、RAPD、SSR、AFLP和RFLP等研究提供高质量的DNA摸板。 相似文献