Common pathogenetic mechanism for three tumor types in bilateral acoustic neurofibromatosis

Bilateral acoustic neurofibromatosis (BANF) is a genetic defect associated with multiple tumors of neural crest origin. Specific loss of alleles from chromosome 22 was detected with polymorphic DNA markers in two acoustic neuromas, two neurofibromas, and one meningioma from BANF patients. This indicates a common pathogenetic mechanism for all three tumor types. The two neurofibromas were among three taken from the same patient, and both showed loss of identical alleles demonstrating that the same chromosome suffered deletion in both tumors. The third neurofibroma from this patient showed no detectable loss of heterozygosity, which suggests the possibility of a more subtle mutational event that affects chromosome 22. In the two acoustic neuromas, only a portion of chromosome 22 was deleted, narrowing the possible chromosomal location of the gene that causes BANF to the region distal to the D22S9 locus in band 22q11. The identification of progressively smaller deletions on chromosome 22 in these tumor types may well provide a means to clone and characterize the defect.     

Chromosome 17 deletions and p53 gene mutations in colorectal carcinomas

Previous studies have demonstrated that allelic deletions of the short arm of chromosome 17 occur in over 75% of colorectal carcinomas. Twenty chromosome 17p markers were used to localize the common region of deletion in these tumors to a region contained within bands 17p12 to 17p13.3. This region contains the gene for the transformation-associated protein p53. Southern and Northern blot hybridization experiments provided no evidence for gross alterations of the p53 gene or surrounding sequences. As a more rigorous test of the possibility that p53 was a target of the deletions, the p53 coding regions from two tumors were analyzed; these two tumors, like most colorectal carcinomas, had allelic deletions of chromosome 17p and expressed considerable amounts of p53 messenger RNA from the remaining allele. The remaining p53 allele was mutated in both tumors, with an alanine substituted for valine at codon 143 of one tumor and a histidine substituted for arginine at codon 175 of the second tumor. Both mutations occurred in a highly conserved region of the p53 gene that was previously found to be mutated in murine p53 oncogenes. The data suggest that p53 gene mutations may be involved in colorectal neoplasia, perhaps through inactivation of a tumor suppressor function of the wild-type p53 gene.     

Concerted nonsyntenic allelic loss in human colorectal carcinoma

Familial polyposis coli (FPC) is caused by an autosomal dominant gene on chromosome 5, and it has been proposed that colorectal cancer in the general population arises from loss or inactivation of the FPC gene, analogous to recessive tumor genes in retinoblastoma and Wilms' tumor. Since allelic loss can be erroneously scored in nonhomogeneous samples, tumor cell populations were first microdissected from 24 colorectal carcinomas, an additional nine cancers were engrafted in nude mice, and nuclei were flow-sorted from an additional two. Of 31 cancers informative for chromosome 5 markers, only 6 (19%) showed loss of heterozygosity of chromosome 5 alleles, compared to 19 of 34 (56%) on chromosome 17, and 17 of 33 (52%) on chromosome 18. Therefore, it appears that (i) FPC is a true dominant for adenomatosis but not a common recessive gene for colon cancer; and (ii) simple Mendelian models involving loss of alleles at a single locus may be inappropriate for understanding common human solid tumors.     

镉胁迫对青菜幼苗某些生理特性以及基因组多态性的影响

以青菜品种"苏州青"为试材,采用生理生化方法和随机扩增多态性DNA(RAPD)技术,研究了镉(Cd)胁迫对青菜幼苗相关生理特性以及基因组多态性的影响。结果表明:不同浓度(1、5、10、20、40 mg·L-1)的Cd处理20 d后,青菜幼苗的鲜重、根伸长受到显著抑制,叶片的叶绿素以及蛋白质含量下降,丙二醛含量和细胞膜透性增加。选用8条寡核苷酸引物(10 bp)对青菜幼苗基因组DNA进行RAPD扩增,对照组的RAPD图谱中可分辨出55条谱带,Cd胁迫后的RAPD图谱发生改变,呈现谱带的增加、缺失和荧光强度的改变,并且与Cd浓度剂量-效应相关。随着Cd浓度的增加,幼苗叶片基因组的模板稳定性逐步下降,并且RAPD多态性与青菜幼苗的生理指标具有显著的相关性。以上结果表明,利用RAPD技术获得的DNA多态性变化谱带可作为检测Cd对青菜毒性效应的生物标记物。     

Alterations of myc, myb, and rasHa proto-oncogenes in cancers are frequent and show clinical correlation

Alterations of c-myc, c-rasHa, or c-myb oncogenes were found in more than one-third of human solid tumors. Amplification of c-myc occurred in advanced, widespread tumors or in aggressive primary tumors. Apparent allelic deletions of c-rasHa and c-myb can be correlated with progression and metastasis of carcinomas and sarcomas.     

RFLP缩减杂交技术的改良与水稻雄性不育细胞质基因组差异片段的克隆

对基因组RFLP缩减杂交技术进行了改良,即用连接成大分子的Driver DNA与Tester DNA杂交、再利用PCR纯化试剂盒对大片段DNA回收率低的原理去除大部分Driver DNA以及与之杂交的非特异性Tester DNA片段.经过PCR扩增和克隆,获得多态性片段,可快速获得在亲本间有差异的分子标记．用该技术对水稻野败型雄性不育细胞质和正常细胞质DNA进行RFLP缩减杂交,获得了2个在不育系细胞质DNA与保持系细胞质DNA间的差异片段。     

基于转录组测序开发糜子SSR标记

【目的】 以6份不同地理来源的糜子资源为试验材料,基于前期转录组测序获得的1 000对SSR引物,选出200对进行多态性检测,以期构建一批可以准确评估糜子种质遗传差异的分子标记。【方法】 用Primer Premier 5.0软件设计引物,改良CTAB法提取DNA,PCR扩增DNA和聚丙烯酰胺凝胶电泳筛选引物多态性;用PowerMarker 3.25和PopGen 1.32计算遗传多样性参数。【结果】 200对引物中97对呈单态性,80对呈多态性。单碱基序列重复引物有20对,10对具多态性,其重复基元是A（50%）和T（50%）。二碱基序列重复引物有36对,15对具多态性,其碱基重复类型有7种（AG最多,TC、GC和GA次之,CA、TA和AC最少）。三碱基序列重复引物有144对,55对具多态性,其碱基重复类型有24种（GGC、GCG和GCC最多,GAA、GCT和CGC等次之,ACC、AGG、CAG、CGT、AAG、AAC、TCG、CGA、ATT、CAA和CCA最少）。就引物分辨率(Rp)值而言,1—3碱基序列重复引物分别为0.67—4.67（平均2.07）、1.33—4.33（平均2.73）和0.67—4.00（平均1.83）。基于Rp值分析SSR分布频次,发现80个标记分布在5个区间：0—1、1—2、2—3、3—4和4—5,分别包含17（21.25%）、36（45.00%）、11（13.75%）、14（17.50%）和2个（2.50%）。就$\overline{\text{Rp}}$值而言,1—3碱基序列重复引物分别为0.33—0.67（平均0.51）、0.40—0.78（平均0.59）和0.33—0.83（平均0.59）。单碱基序列重复标记共检测到22个等位变异,每个位点为2—3个（平均2.2000）,其中8个和2个位点分别具2和3个变异;二碱基序列重复标记共检测到38个等位变异,每个位点为2—3个（平均2.5333）,其中7个和8个位点分别具2和3个变异;三碱基重复标记共检测到136个等位变异,每个位点为2—3个（平均2.4727）,其中29个和26个位点分别具2和3个变异。就多态性信息含量而言,1—3碱基序列重复引物分别为0.3750—0.5355（平均0.4293）、0.2392—0.7438（平均0.4293）和0.2392—0.7438（平均0.3989）。就多样性指数而言,1—3碱基序列重复分别为0.6365—1.0776（平均0.7497）、0.5623—1.0986（平均0.8339）和0.5623—1.0889（平均0.8312）。【结论】 基于转录组测序结果,检测200对SSR引物的多态性,发现177对（88.5%）可以扩增出完整条带,其中80对具多态性,多态率为40%。     

Structure of Allozymatic Diversity of Ten Temperate and Adapted Exotic Breeding Populations of Maize (Zea mays L.)

Ten temperate and adapted exotic breeding populations of maize were studied with electrophoretic techniques. Three isozyme systems coded by nine allozyme loci were used for evaluating the genetic variability within and among populations.The results revealed that 78.57% of allozyme loci were polymorphic. Low allelic variation with a mean number of 1.84 alleles per locus per population was detected. But, these populations still maintained higher level of heterozygosity;moreover, the exotic populations had greater gene diversity than the temperate populations. All the populations were non-panmictic with negative Wright's fixation indexes (-0.091- -0.424). The tropical BS16 was typified by maximum allelic richness, percent of polymorphic loci and heterozygosity. More than 93% of the gene diversity maintained within populations, and the genetic differentiation among populations was low (0.002-0.191). Multivariate analysis demonstrated that the tropical BS29 diverged from other populations in the reverse direction. The temperate BS9 and tropical BS 16 were divergent each other, and highly differentiated from other temperate and tropical populations, consequently, these two populations would be analogically postulated as potential germplasms to establish new heterotic groups for temperate maize breeding programs.     

Success and virulence in Toxoplasma as the result of sexual recombination between two distinct ancestries

Toxoplasma gondii is a common human pathogen causing serious, even fatal, disease in the developing fetus and in immunocompromised patients. Despite its ability to reproduce sexually and its broad geographic and host range, Toxoplasma has a clonal population structure comprised principally of three lines. We have analyzed 15 polymorphic loci in the archetypal type I, II, and III strains and found that polymorphism was limited to, at most, two rather than three allelic classes and no polymorphism was detected between alleles in strains of a given type. Multilocus analysis of 10 nonarchetypal isolates likewise clustered the vast majority of alleles into the same two distinct ancestries. These data strongly suggest that the currently predominant genotypes exist as a pandemic outbreak from a genetic mixing of two discrete ancestral lines. To determine if such mixing could lead to the extreme virulence observed for some strains, we examined the F(1) progeny of a cross between a type II and III strain, both of which are relatively avirulent in mice. Among the progeny were recombinants that were at least 3 logs more virulent than either parent. Thus, sexual recombination, by combining polymorphisms in two distinct and competing clonal lines, can be a powerful force driving the natural evolution of virulence in this highly successful pathogen.     

An X chromosome gene, WTX, is commonly inactivated in Wilms tumor

Wilms tumor is a pediatric kidney cancer associated with inactivation of the WT1 tumor-suppressor gene in 5 to 10% of cases. Using a high-resolution screen for DNA copy-number alterations in Wilms tumor, we identified somatic deletions targeting a previously uncharacterized gene on the X chromosome. This gene, which we call WTX, is inactivated in approximately one-third of Wilms tumors (15 of 51 tumors). Tumors with mutations in WTX lack WT1 mutations, and both genes share a restricted temporal and spatial expression pattern in normal renal precursors. In contrast to biallelic inactivation of autosomal tumor-suppressor genes, WTX is inactivated by a monoallelic "single-hit" event targeting the single X chromosome in tumors from males and the active X chromosome in tumors from females.     

花生优异种质的分子标记与遗传多样性分析

【目的】通过对花生遗传多样性的研究,为花生育种提供理论依据。【方法】运用ISSR和SRAP2种标记对24份重要花生种质资源的遗传多样性进行分析。【结果】32条ISSR引物中的13条引物共扩增出390条条带,其中多态性条带有293条,75.13%的条带可以揭示材料之间的遗传差异;252对SRAP引物中有229对引物为有效引物,共扩增出5827条可读的条带,其中多态性条带为3966条,平均每对引物可扩增17.32条条带。利用2种分子标记计算的相似系数的变化范围为0.60—0.80,将24份种质按系统聚类分析可以分为7组,按主坐标分析可以分为8组,从分子水平上解释了这些种质资源的遗传多样性水平和亲缘关系。【结论】ISSR和SRAP是非常有效、稳定和可靠的分子标记,可为花生育种的亲本利用及遗传连锁图的构建提供重要的科学依据。     

不同核桃品种的SSR分析

应用SSR技术对21个核桃品种和1个枫杨品种的基因组DNA进行遗传多样性的研究。用筛选出的10对引物对供试材料进行SSR分析,共扩增出67条清晰的谱带,不同引物扩增出的DNA条带数量在4～24条之间,分子量大小在85～300 bp之间,依据特征谱带,可直接鉴别出供试品种,其中引物WGA32的鉴别效率最高。对扩增出的67条带进行聚类分析,结果表明,普通核桃和枫杨亲缘关系较远,不同地域起源的品种界限不明显。     

Development of Soybean EST-SSR Markers and Their Use to Assess Genetic Diversity in the Subgenus Soja

Developing expressed sequence tag-derived SSR （EST-SSR） markers is imperative in genetic research. In this paper, we reported 37 EST-SSR markers which were developed from 286 unigenes obtained from soybean cDNA library. Among the 286 markers designed for the 4 accessions of Glycine max and 6 of its wild progenitor （G. soja） within the subgenus Soja, 209 markers amplified DNA fragments, taking 73.1% and 37 markers appeared to be polymorphic, which was 12.9% of the total. The 37 loci detected a total of 142 alleles, while the PIC values varied from 0.194 to 0.794. Both the number of alleles per locus and PIC value were significantly related to the SSR motif. Six EST-SSR loci may be fixed for different alleles between G. max and G. soja since they were particularly polymorphic among the 6 G. soja accessions. A neighbor-joining tree placed the G. max accessions together as a group within the G. soja, though the average genetic distance among G. soja accessions was much higher. These new EST-SSRs markers will be useful for genetic diversity analysis, genetic mapping construction and gene discovery in Soja subgenus.     

Structure of AIIozymatic Diversity of Ten Temperate and Adapted Exotic Breeding Populations of Maize （Zea mays L.）

Ten temperate and adapted exotic breeding populations of maize were studied with electrophoretic techniques. Three isozyme systems coded by nine allozyme loci were used for evaluating the genetic variability within and among populations. The results revealed that 78.57% of allozyme loci were polymorphic. Low allelic variation with a mean number of 1.84 alleles per locus per population was detected. But, these populations still maintained higher level of heterozygosity; moreover, the exotic populations had greater gene diversity than the temperate populations. All the populations were non-panmictic with negative Wright＇s fixation indexes （-0.091— -0.424）. The tropical BS 16 was typified by maximum allelic richness, percent of polymorphic loci and heterozygosity. More than 93% of the gene diversity maintained within populations, and the genetic differentiation among populations was low （0.002—0.191）. Multivariate analysis demonstrated that the tropical BS29 diverged from other populations in the reverse direction. The temperate BS9 and tropical BS16 were divergent each other, and highly differentiated from other temperate and tropical populations, consequently, these two populations would be analogically postulated as potential germplasms to establish new heterotic groups for temperate maize breeding programs.     

藏绵羊DQA1基因多态性分析

 【目的】研究藏绵羊DQA1基因多态性,确定其等位基因数、核苷酸多态位点、氨基酸多态位点及各等位基因间的遗传关系,同时分析其进化意义。【方法】采用PCR-SSCP方法检测了900只藏绵羊DQA1基因第2外显子多态性;克隆、测序群体内变异产生的各等位基因序列,并分析序列数据。【结果】发现了17个DQA1的等位基因,包括缺失的1种基因,其中5个为发现的新等位基因。16个单倍型序列中发现56个核苷酸多态位点,27个氨基酸多态位点。【结论】藏绵羊DQA1基因第2外显子具有丰富的多态性,群体中可能蕴藏着更多的遗传资源;藏绵羊DQA1基因最初可能是由2个等位基因突变分化成两大类等位基因的;藏绵羊DQA1基因第2外显子序列与牛的DQA1基因第2外显子序列具有较高的同源性,预示绵羊和牛的DQA1基因最早可能来源于它们分歧以前的共同祖先原始序列;DQA1基因在与其相关的特定抗原刺激下发生的免疫应答反应在绵羊和牛上具有相似性;新等位基因C的139位发现了1个新的核苷酸突变位点（A/G）,属同义突变;5个新发现的DQA1等位基因遗传关系较近,可能由同一等位基因突变产生。     

基于荧光检测技术的多花黑麦草EST-SSR指纹图谱的构建

【目的】构建基于EST-SSR荧光标记的多花黑麦草(Lolium multiflorumLam.)DNA指纹鉴定体系,为多花黑麦草品种鉴定提供高通量技术手段,为不同品种的合理应用提供参考依据,有效保护农民利益和育种权益。【方法】利用表型性状差异大的3个品种(特高Tetragold、长江2号Changjiang No.2和阿伯德Aubade),通过聚丙烯酰胺凝胶电泳从200对EST-SSR引物中筛选出扩增条带清晰、多态性好、扩增稳定的30对引物。在筛选出的每对引物5′端添加荧光标记FAM后,采用毛细管法通过DNA分析仪检测200个单株不同等位变异的扩增片段,从30对EST-SSR引物中筛选出25对扩增稳定的荧光引物,建立基于高通量荧光SSR标记的多花黑麦草品种鉴定体系。【结果】通过25对EST-SSR引物构建的DNA指纹图谱来进行10个多花黑麦草材料的品种鉴定。25对EST-SSR引物共检测到127个等位基因,等位变异扩增片段长度范围为51—249 bp,每对引物可检测到有效等位基因数为2—11个,特异等位基因最多可检测到11个(N101),平均每对引物4.00个;多态性位点的比率范围为33.33%—100.00%。平均PIC值为0.702,Shannon指数最大为3.322(N101),平均为1.929,基因多样性指数变幅为0.159—0.500,平均0.318,可鉴别的材料数为0—10个;其中14对特征引物在10个品种(系)上可检测出25个特异等位基因。综合来看,引物N101在200对引物中鉴别效率最高,可直接将10个多花黑麦草品种(系)区分开,在长江2号、赣选1号和川农2号上同时检测出特异等位基因。但由于多花黑麦草在品种间与品种内变异均较高,因此为鉴定更多材料,从25对引物中选择了6对扩增和检测效果较好的引物(N54、N101、N146、N151、N154、N156),6对引物可检测到的稳定等位基因数均不小于19个,在达伯瑞和邦德上最多可检测到22个等位基因。通过6对高效引物构建了10个多花黑麦草品种(系)的DNA指纹图谱,包括标准模式图、图谱代码和图谱QR编码。首次利用EST-SSR荧光标记毛细管电泳检测,为10个多花黑麦草材料分别构建了唯一的指纹代码和QR编码。【结论】利用6对高效引物构建了多花黑麦草SSR高通量鉴定体系,其中荧光引物N101多态性最高,可直接鉴别10个多花黑麦草品种(系)。     

茄子主要栽培种遗传多样性分析

为了明确我国茄子主要栽培种的遗传背景,对主要来自广西及全国部分地区的54份茄子栽培种进行表形标记和ISSR标记分析.结果表明,在37个表形性状中,表现为多态性有33个,占89.18％;从50条ISSR引物中共筛选出8条多态性明显、条带清晰、反应稳定的引物,扩增出68条谱带,平均每条引物扩增出8.5条带,多态性带58条,占85.29％;品种间两类型标记遗传相似系数均在0.04 ～1.00之间,总体平均数分别为0.51、0.60,表明个别品种间遗传差异很大,但群体整体遗传基础相对较狭窄;应用SPSS软件聚类分析,两类标记均能在欧氏距离14～15间将54个栽培种划分为6个类群,其中表形标记类群的划分与地理来源有一定关系,而分子标记类群的划分与地理来源没有明显的联系.     

不同地理种源雷公藤的RAPD分析

应用RAPD技术,对25个不同地理种源的雷公藤进行了多态性分析。采用POP-GENE32软件对25个种源进行聚类分析,DPS(9.5)软件对所有个体进行聚类分析,并用UPGMA法对扩增DNA片段数据构建系统发育树。结果表明:多态性条带百分率为91.14%,Shannon's信息指数I=0.4472,Nei's基因多样性指数H=0.2973,观测等位基因数Na=1.911 4,有效等位基因数Ne=1.5120。这表明25个种源具有较高的遗传多样性水平,因此收集不同种源以保存尽可能多的遗传多样对于雷公藤的良种选育是非常有必要的。