BALB／C小鼠的白内障突变与大片段小卫星DNA多态性的变化

为了探讨ＢＡＬＢ／Ｃ小鼠白内障突变品系与ＢＡＬＢ／Ｃ小鼠保种品系二者间小卫星ＤＮＡ多态性的变化,为进一步研究变异小卫星位点与白内障突变基因的关系最终找到导致这种白内障形成的基因奠定基础,采用α－＾３２Ｐ－ｄＣＴＰ标记的人类小卫星ＤＮＡ探针Ｍｙｏ,通过Ｓｏｕｔｈｅｒｎ杂交比较了正常ＢＡＬＢ／Ｃ和ＢＡＬＢ／Ｃ白内障突变体小卫星ＤＮＡ多态性的变化。结果：ＢＡＬＢ／Ｃ白内障小鼠种内ＤＮＡ多态性带纹分布高度     

四个探针产生的家禽 DNA 指纹图谱

对四个多位点小卫星探针 33.6,33.15,α珠蛋白-3’HVR 和 M13用于家禽的DNA 指纹分析的可行性进行了探讨,较详细地报导了用不同探针在鸡、鸭、鹌鹑上产生 DNA 指纹图的方法.结果表明,用我们所采用的方法,四个探针都能在鸡、鸭、鹌鹑上产生信息量大、分辨率较高的 DNA 指纹图.     

鸡的小卫星探针产生的猪DNA指纹图

以一个鸡的小卫星cMS18为探针,对五指山小型猪、长白猪及枫泾猪进行了DNA指纹分析。结果表明,五指山小型猪的相似系数(0.721)显著高于长白猪(0.503)和枫泾猪(0.484),在遗传背景上也与后两者相距甚远。家系分析表明, 鸡的小卫星探针产生的猪的DNA指纹图的遗传符合孟德尔规律。Abstract:A chicken minisatellite probe cMS18 was used to generate DNA fingerprints of Wuzhisshan microping,Landrace pig and Fengjing pig.It was revealed that similarity coefficient of Wuzhishan microping(0.721)was much higher than both that of Landrace pig(0.503)and of Fengjing pig(0.484).It was shown that Wuzhishan micropig was genetically distant to Landrace pig and Fengjing pig.Analysis of a porcine pedigree using cMS18 probe manifested that the segregation of DNA fingerprint bands was consistent with the Mendelian fashion.     

DNA探针的诊断应用

<正>使用重组DNA技术制备特异的分子(常为DNA)探针已为医学和兽医诊断实验室提高对传染病、遗传紊乱、恶性肿瘤的诊断提供了新的强有力的工具,同时也给完成如组织分型、亲缘关系试验这类工作提供了更灵敏感更特异的方法,该技术对法医学也是很有用的(如表1)。DNA探针作为诊断试剂的应用或使实验室减少周转时间,拓宽所检测、鉴别和/或定量的试样的范围,且由于简化     

用随机扩增多态性DNA产物做探针产生鸡的DNA指纹图

我们用12个随机扩增多态性DNA(RAPD)引物对来自不同品系的4只鸡进行了RAPD分析,在扩增出的共99条带中,表现多态性的带为38条,占总带数的38％.回收了4个表现个体特异性的RAPD产物,当用鸡的基因组总DNA探针与它们杂交时,其中3个表现阳性,说明RAPD方法扩增出的高变异产物含有重复序列.用含重复序列的个体特异性RAPD产物作探针,与无关个体鸡基因组DNA的HaeⅢ酶切产物进行DNA印迹,获得了变异性较高的DNA指纹图谱.因此,高变异的RAPD产物可以有效地用作DNA指纹探针.     

人巨细胞病毒的生物素DNA探针的制备和应用

本研究用克隆的HCMV AD169株DNA片段,制备了生物素标记的DNA探针,建立了检测临床脐带血、尿标本中HCMV DNA的核酸探针杂交方法。该探针可测出100pg同源DNA,不与人胚肺细胞、Hep-2细胞DNA以及其他疱疹病毒的DNA发生反应。用核酸杂交方法检测了30份脐带血标本,有11例阳性,阳性率为33％。10例孕妇尿标本中,3例阳性,阳性率为30％。检测结果表明：我们建立的生物素标记的HCMV DNA探针的点杂交法,具有高度的特异性、敏感性,比分离病毒法更迅速,可用于HCMV感染的临床标本的病毒核酸检测。     

植物特异性DNA探针的制备与应用研究进展

本文简述了植物特异性DNA探针的种类,介绍了特异性探针的制备方法,主要是特异性DNA序列的分离与筛选,总结了植物特异性DNA探针在种质鉴定、外源染色体识别、核型分析和基因组研究等方面的应用,提出了存在的问题与应用前景。Abstract:In this paper,the types of specific DNA probes from plant were briefly described,and some methods of constructing specific probes were introduced,mainly were the isolation and screening of specific DNA sequences.The applications of the specific probes in the germplasm and foreign chromosome identifications,the karyotype and genome analyses were also summarized and discussed.     

Cloning, quantification and characterization of a minisatellite DNA sequence from common bean Phaseolus vulgaris L.

 We describe the cloning and the characterization of a 130-bp DNA fragment, called OPG9-130, amplified from bean (Phaseolus vulgaris L.) genomic DNA. This fragment corresponds to a minisatellite DNA sequence containing seven repeats of 15 bp which differ slightly from each other in their sequence. Southern analysis showed that the core sequence of 15 bp is repeated in clusters dispersed throughout the genome. The use of this fragment as a probe allowed us to identify common bean lines by their DNA fingerprints. We suggest that OPG9-130 will be useful for line identification as well as for the analysis of genetic relatedness between bean species and lines. Received: 14 February 1998 / Accepted: 10 February 1998     

Minisatellite DNA fingerprints of salmonid fishes

The human minisatellite probes 33.6 and 33.15 cross-hybridized to DNA digests of Atlantic salmon, brown trout and rainbow trout revealing complex multi-banded patterns. These DNA fingerprints (in excess of 40 resolvable fragments in some cases) were highly polymorphic, individual specific and found to be stable, both somatically and in the germline. Pedigree analysis of an Atlantic salmon family confirmed that the minisatellite fragments showed Mendelian inheritance. With only a single occurrence of linkage and allelism being observed it is likely the minisatellite loci are widely distributed throughout the salmonid genome. The potential applications for both multi- and single locus minisatellite probes in salmonid research are discussed.     

Minisatellite DNA fingerprints of Atlantic cod (Gadus morhua)

The human minisatellite probes 33.6 and 33.15 cross–hybridized to Hae III and Hinf I digested cod DNA, revealing complex fragment patterns in both Arctic and coastal cod. The DNA fingerprints were highly polymorphic, individual specific and stable in the germline. The potential applications of multi locus probes in cod research are discussed.     

Minisatellite germline mutation rate in the Techa River population

Germline mutation at eight minisatellite loci has been studied among the irradiated families from the Techa River population and non-exposed families from the rural area of the Chelyabinsk and Kurgan Oblasts. The groups were matched by ethnicity, parental age, occupation and smoking habit. A statistically significant 1.7-fold increase in mutation rate was found in the germline of irradiated fathers, whereas maternal germline mutation rate in the exposed families was not elevated. Most of the minisatellite loci showed an elevated paternal mutation rate in the exposed group, indicating a generalised increase in minisatellite germline mutation rate in the Techa River population. These data suggest that the elevated minisatellite mutation rate can be attributed to radioactive exposure. The spectra of paternal mutation seen in the unexposed and exposed families were indistinguishable.     

人乳头瘤病毒芯片寡核苷酸探针的研究

高危型人乳头瘤病毒（Human papillomavims,HPV）是宫颈癌的主要致病因子。利用Arraydesigner2．0和BLAST等生物学软件对10种型别的人乳头瘤病毒全基因组序列进行分析,设计高特异性、熔解温度（Tm）和GC含量相近的60mer HPV型特异性寡核苷酸探针,用于HPV检测芯片的制备,并对其中四型最常见HPV病毒（HPV6,11,16,18）探针的有效性进行初步验证,结果表明设计所得的探针型特异性好,可以应用于HPV的检测与分型。     

Minisatellite MS32 Alleles Show Population Specificity Among Thai,Chinese, and Japanese

Lineages of structurally related alleles at minisatellite MS32 in human populations show considerable differentiation at the continental level. However, the regional specificity of these lineages remains unknown. We now describe the comparison of allele structures in Thai, Han Chinese, and Japanese populations with lineages previously established for North Europeans and Africans. The great majority of alignable Asian alleles showed their closest structural relative in Asia, with few instances of preferential alignment of Asian with European alleles and only one isolated incident showing a best match with an African allele. Further, there was a strong tendency, most marked for Japanese, for Asian alleles to align preferentially with other alleles from the same population, indicating strong regional specificity of allele lineages. This rapidly evolving minisatellite can therefore serve as a lineage marker for exploring recent events in human population history and dissecting population structure at the fine-scale level, as well as being an extremely informative DNA marker for personal identification.     

Minisatellite DNA markers in the chicken genome

This paper reports the detailed characterization of multilocus minisatellite DNA fingerprints in the chicken. Results are presented of DNA fingerprint segregation analyses carried out in three chicken pedigrees, calculating the number of detected loci, testing for Mendelian inheritance, and cosegregation among fingerprint bands. Two pedigrees (families 1 and 2) were analysed using the Jeffreys probes 33.6 and 33.15 only, and one pedigree (family 3) was analysed using 33.6, 33.15. 3′α-globin HVR and M13 protein III gene repeat. Mean band transmission frequencies in families 1 and 2 were near to the Mendelian expectation of 0.5 and no mutations were observed. Family 3 showed transmission frequencies slightly exceeding 0.5. Linkage among bands was higher than observed in some other avian species, with each allele represented by a mean of 1.48 HaeIII fragments. Cosegregation of heterozygous parental fragments representing distinguishable loci followed the expected binomial distribution. The number of minisatellites detectable by the four probes was estimated to be 217. The pattern of cosegregation among those minisatellite loci was tested against that expected for different levels of recombination through the use of a simulation model. We conclude that most minisatellites are unlinked and probably widely dispersed in the chicken genome.     

Minisatellite polymorphisms of the SLC6A19: susceptibility in hypertension

The SLC6A19 is a human homolog of B⁰AT1 that encodes a neutral amino acid transporter. We examined the distribution of VNTR (variable number of tandem repeats; minisatellites) and conducted polymorphic analysis of SCL6A19 isolated from the genomic DNA of controls and multi-generational families. The SLC6A19 was found to contain seven blocks of minisatellites, 3 of which (SLC6A19-MS1, -MS4, and -MS7) showed polymorphism and were found to be transmitted through meiosis following Mendelian inheritance in seven families. These minisatellite polymorphisms may be useful markers for paternity mapping and DNA fingerprinting. Furthermore, we conducted a case-control study in which genomic DNA from 400 controls and 205 cases with essential hypertension was compared. A statistically significant association was identified between rare SLC6A19-MS7 alleles and the occurrence of hypertension (odds ratio, 7.87; 95% confidence interval, 0.88-70.66; and p = 0.028). These findings suggest that the rare SLC6A19-MS7 allele may be a risk factor for hypertension.     

小鼠DNA的限制性片段长度多态性分析

用人类肌红蛋白基因内含子中３３ｂｐ的小卫星ＤＮＡ重复序列为探针,经ｐＢＲ３２２的ＥｃｏＲ Ｉ正向测序引物和α－３２Ｐ－ｄＣＴＰ在Ｔａｑ ＤＮＡ聚合酶作用下做合成标记,与６种小鼠的ＤＮＡ限制性片段杂交。结果显示,每个品系平均出现可分辨带纹在１３条以上,不同品系小鼠间的杂交带纹表现出高度的多态性。多态性带纹主要分布在６￣２３ｋｂ区段;同一近交系的不同个体间的带纹特点及分布完全一致;ＣＳ７与ＡＫＲ及二者     

Oligonucleotide probes containing pyrimidine analogs reveal diminished hydrogen bonding capacity of the DNA adduct O6-methyl-G in DNA duplexes

Oligonucleotide hybridization probes containing nucleoside analogs offer a potential strategy for binding specific DNA sequences that bear pro-mutagenic O⁶-G alkylation adducts. To optimize O⁶-Me-G-targeting probes, an understanding of how base pairs with O⁶-Me-G are stabilized is needed. In this study, we compared the ability of O⁶-Me-G and G to hydrogen bond with three pyrimidine-like nucleobases (Z, 4-thio-U, and 3-deaza-C) bearing varied hydrogen bond donor and acceptor groups. We found that duplexes containing the pyrimidine analog nucleoside:G pairs were more thermodynamically stable than those containing pyrimidine analog nucleoside:O⁶-alkyl-G pairs. Thus, hydrogen bonding alone was not sufficient to impart selectivity to probes that target O⁶-G alkylation adducts in DNA.