共查询到19条相似文献,搜索用时 218 毫秒
1.
应用Bulked-DNA寻找白菜型油菜核雄性不育基因的RAPD标记 总被引:8,自引:2,他引:6
采用随机扩增多态DNA(RAPD)标记方法从白菜型油菜核不育两用系中筛选出了一个与白菜型 油菜育性基因连锁的RAPD标记。该DNA片段大小约0.72kb,与育性基因之间的遗传连锁距离为6.08cM,LOD值为9.10。 相似文献
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斜纹夜蛾核多角体病毒egt基因的克隆和部分序列分析 总被引:7,自引:0,他引:7
闫庆生 《中山大学学报(自然科学版)》1998,37(3):125-127
用AcMNPVegt基因中的保守区部分片段为探针,通过Southern杂交确定SlMN-PVegt基因的位置在PstⅠ4970bp和XbaⅠ2600bp的片段上.采用双链DNA序列分析方法测序,在2600bp片段上的EcoRⅠ位点两侧得到594bpDNA碱基序列,用计算机DNASIS和PROSIS软件,将594bpDNA碱基序列所推测的氨基酸序列与AcMNPV和SliMNPV的egt基因氨基酸序列进行同源比较,其同源性分别为45%和86.5%. 相似文献
3.
斜纹夜蛾NPV多角体基因的克隆和部分测序 总被引:1,自引:1,他引:1
本文对SINPV基因组作了酶解分析,测得其基因组大小为145kb,并用双酶法确定了SINPV基因组的HindⅢ和PstⅠ物理图谱。以含AcNPV多角体基因的质粒pAC-Ⅰ的SalI-C片段为探针,对SINPVDNA酶切片段southern转印杂交结果,初步判断多角体基因定位于PstI-B/C/D片段、BglⅡ-C/D片段、BamHI-B/C片段和EcoRI-A/B片段上,且SINPV与AcNPV多角体蛋白基因有64%的同源性,而以大肠仟菌质粒pUC19为载体对SINPV的多角体基因试克隆,得到带有BglⅡ-PstⅠ双酶切片段的2个克隆子。对这两个杂交阳性克隆子之一的核苷酸序列测定,表明插入片段与BmNPV多角体基因上游序列亦有一定同源性。 相似文献
4.
水稻共生菌DX01的分子鉴定及电击转化条件的初步探索 总被引:4,自引:1,他引:3
用PCR的方法扩增了水稻表面共生菌DX01的16S rDNA片段,并进行了部分序列的测定和BLAST同源序列比较分析,结果表明,该序列3’端550bp与短小芽孢杆菌(Bacillus pumilus)16 S rDNA的同源性达99%,5‘端680bp为98%,故DX01应为B.pumilus。对其电击法转化的部分条件进行了初步的探索,结果表明:当细菌处于D(600)为0.9的生长期时,使用PEB 相似文献
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维甲酸激活表达的人肺腺癌细胞分化相关基因的筛选 总被引:1,自引:0,他引:1
通过对维甲酸诱导的人肺腺癌细胞GLC-82 cDNA消减文库的筛选,获得2个特异存在于维甲酸诱导1天cDNA文库中的cDNA片段RA97及RA107,测序后与最新GenBank提供的序列比较,RA97与人22号染色体臂13区PAC408N23同源性很强,其中一段与PAC408N23完全同源,此片段与可能的肿瘤抑制基因SNC6同源性也相当强,其中一段与SNC6完全同源,提示这一cDNA片段可能与肿瘤 相似文献
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pANGPD是用构巢曲霉GPD基因片段为探针,从黑曲霉基因组文库中筛选到的一个三磷酸甘油醛脱氢酶基因(GPD)阳性克隆.作者对该阳性克隆进行了亚克隆和序列测定,结果表明,基因片段总长2441个核苷酸(n.t),其中5’-端非编码区长981n.t,编码区(从起始密码子ATG到终止密码子TAG)长1446n.t,3’-端非编码区长24n.t,基因启动子区含有gpd-盒和CT-盒序列,但无明显的CAAT盒和TATA盒序列.GPD基因由7个外显子和7个内含子构成,外显子全长1008n.t,编码的蛋白质长336个氨基酸残基.通过序列比较分析,发现该基因与构巢曲霉的GPD基因有许多相似之处;它们的基因启动子序列都含有gpd-盒和CT-盒,其同源性分别为96%和65%;推测的两个GPD蛋白的氨基酸同源性高达90%;两个基因所含内含子数目及位置相同. 相似文献
8.
斜纹夜蛾NPVp74基因的克隆及部分序列分析 总被引:6,自引:0,他引:6
以含p74基因的AcNPVEcoRⅠ-P片段作探针,与SlNPV基因组在非严格条件下进行Southern杂交,将SlNPVp74基因定位于4900bp的SlNPVXhoⅠ-P片段.对SlNPVXhoⅠ-P片段中的一段940bp的DNA片段进行序列测定,通过internet经BLAST与Gen-Bank中的database比较分析发现,序列中的一段243nt序列与AcNPVp74基因有60%的同源性,一段81nt序列与AcNPVp74基因的同源性高达79%.与CfNPVp74相比,2段264nt和100nt序列分别有60%和71%的同源性.测得序列中的部分序列与BmNPV、OpNPVp74基因也有高达58%~78%的同源性.同时,这部分序列编码的氨基酸与AcNPV及OpNPVP74蛋白的氨基酸序列也有很高的同源性.结果表明所测序列的一部分为SlNPVP74蛋白C-端的编码序列. 相似文献
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籼型光敏核不育水稻的RAPD分析 总被引:2,自引:0,他引:2
以籼型光敏核不育水稻8902s及其可育近等基因系材料8902的核DNA为模板,进行RAPD分析,从检测过的15个引物中发现引物OPA-04在可育品系和不育品系中扩增出1个1000bp的差异带,并初步认为此差异与育性相关 相似文献
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探讨克服肿瘤细胞多药耐药(MCR1)性的方法,提高化疗效。本采用多药耐药反义基因(MDR1-RSPS-ODN)逆转K562/ADM肿瘤细胞的MDR1,诱导肿瘤细胞凋亡MDR1-AS PS-ODN诱导K562/ADM细胞株细胞产生大量DNA断片,FACS检测发现几乎全部MRD^+K562/ADM细胞发生凋亡。其结果表明DMDR1-AS PS-ODN能有效、特异地抑制MDR1基因表达,逆转肿瘤细胞的 相似文献
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水稻光温敏雄性不育基因连锁标记的分离与鉴定 总被引:2,自引:0,他引:2
两系法杂交稻是一种利用水稻杂种优势的重要途径,主要依据水稻光温敏雄性不育系在不同条件下的育性转换用于配制杂交种.以培矮64S(PTGMS)与明恢63杂种自交的F2代为供试材料,采用随机放大多态性DNA(RAPD)技术分离与培矮64S的PTGMS基因连锁的分子标记.在F2代2个分别代表可育和不育的群体以及亲本株系中,采用100个RAPD引物扩放基因组DNA筛选多态性DNA片段.RAPD引物S8产生的DNA片段中,除重复顺序外,有一个长度为0.85kb片段为单拷贝.分子杂交表明,这一单拷贝标记与培矮64S的PTGMS基因连锁. 相似文献
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Identification of RAPD marker linked with fertility-restoring gene of cytoplasmic male sterile lines in upland cotton 总被引:5,自引:0,他引:5
Bulked segregant analysis was employed to construct two mixed DNA pools to screen the RAPd marker linked with the fertility-restoring
gene(Rf
i) of upland cotton. A total of 425 arbitrary 10-mer oligonucleotide primers were screened on two DNA pools, bulked male fertile
and sterile DNAs isolated from BC3 segregating population of (0-613-2R X Simian No. 3). Three primers produced repeatable polymorphisms between the paired bulks
and their parents. DNA was extracted and amplified with these three primers for 92 plants of (Zhong 12A-1 × 0-613-2R)F2. Based on the male fertility scoring and RAPD amplification, it is found that one RAPD marker fragment designated OPV-15300 was linked with the fertility-restoring gene (Rf1) with a recombination value of 13.0±2.57%. 相似文献
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Soybean mosaic virus (SMV) is a severe disease in worldwide soybean production. A cross was made between Kefeng No. 1 with
broad spectrum resistance to SMV and Nannong 1138–2, a susceptible cultivar. The inheritance of resistance to SMV strain Sa
prevailing in southern China was analyzed. Results of x2 test from inoculation experiment on parents F1, F2 and F3 lines showed that the resistance to strain Sa was controlled by
a single dominant gene Rsa. BSA method was adopted and 900 random 10-mer primers were used to amplify total DNA from resistant
pool and susceptible pool in order to obtain polymorphic bands in two bulks. 16 primers could generate polymorphic bands,
of which OPW-05 and OPAS-06 could generate the most stable RAPD patterns. RAPD markers OPW-05660 and OPAS-061800 were found to be linked to Rsa. Their order and genetic distance were OPAS-06180022.2cM Rsa10.1 cM OPW-05660. Southern blotting showed that both OPAS-061800 and 0PW-05660 were low copy DNA in genomic DNA. 0PW-05660 has been converted into an RFLP marker successfully. Additionally, pK644H, an RFLP marker, has been identified to be linked
to 0PW-05660, and their genetic distance was 37.4 cM. 相似文献
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丝黑穗病是影响我国高粱生产发展的主要病害之一,在高粱产区每年都有发生,发病率有时高达70 %,给生产造成很大损失,而选育抗病品种是最为有效的抗病策略.分别以高粱2381恢复系(抗病)、矮四恢复系(感病)的叶片为试材,采用CTAB法提取高粱基因组DNA,并应用RAPD筛选技术对高粱基因进行分子标记.在最佳的RAPD反应体系中共筛选了100个RAPD随机引物,筛选出来27个适宜引物,获得了稳定的RAPD扩增结果及多态性较好的DNA谱带.其中有6个引物对DNA扩增产生了差异谱带. 相似文献
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黄瓜全雌性状相关的RAPD分子标记筛选 总被引:1,自引:0,他引:1
用CTAB法提取黄瓜总基因组DNA,采用RAPD技术探寻与黄瓜全雌性相关的分子标记.共筛选39条随机引物,其中有12条引物显示多态性.进一步筛选表明:S10引物能在全雌系黄瓜中扩增出一条约2000 bp的稳定、清晰的特异条带,而在雌雄同株的单株中不存在.经回收、克隆并测序,获得全长2003 bp的序列.在NCBI上进行Blast分析,表明为新发现序列,含有编码62个氨基酸的开放阅读框.其编码短肽的功能尚不确定. 相似文献
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几种番茄品种基因组DNA的相似指数初步分析 总被引:2,自引:1,他引:1
用4种随机引物(10bp),对普通番茄5个品种的基因组DNA进行了PCR扩增,并对其RAPD带谱进行了统计处理,发现不同随机引物扩增出来的RAPD标记所表现的相似指数有所不同,综合考虑,普通番茄种内不同品种同DNA多态性保持了较高的稳定性。 相似文献
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小麦雌性不育基因的微卫星标记定位 总被引:2,自引:0,他引:2
以普通小麦新601×雌性不育小麦XND126的F2群体作为育性调查以及基因标记群体.通过对育性基因的分析,确定在此组合中雌性不育基因由1对主效基因控制;结合混合分组分析法(Bulk Segregant Analysis,BSA),首次对小麦雌性不育基因进行了SSR分子标记,通过对一千对微卫星引物的筛选,确定微卫星引物cfd36标记与主效基因连锁,遗传距离为20.2cM. 相似文献
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Characterization and molecular marker screening of a rice bacteria-resistant gene Xa-min(t) 总被引:3,自引:0,他引:3
To test the resistant spectrum of the Xa-min(t) gene introgressed from Oryza minuta, thirty-four isolates of different bacterial blight pathogen, Xanthomonas oryzae pv. oryzae (Xoo), from 11 countries were used to inoculate the Xa-min(t) introgression line 78-15. Four rice cultivars, IR24, C64 (IRBB21), Nipponbare and Zhonghua 11 were used as controls. The results showed that the Xa-min(t) gene was broad-spectrum and highly resistant to diverse Xoo isolates. The methods of bulk segregant analysis (BSA), randomly amplified polymorphic DNA (RAPD) and sequence characterized amplified regions (SCAR) were used to analyze F2 individuals of the hybrid IR24×78-15 and molecular genetic markers linked to Xa-min(t) gene were identified. A total of 800 arbitrary decamer oligonucleotide primers were used for RAPD analysis. Two RAPD markers, BE05300 and BE061400, produced by primers BE05 and BE06 respectively, were closely linked to the Xa-min(t) gene. Based on the sequences of these two markers, sequence specific primers were designed and used to screen all F2 plants. One RAPD marker, BE05300, was converted into a stable SCAR marker (ScBE05300). Linkage analysis was carried out using markers ScBE05300 and BE061400 on 948 and 719 F2 individuals of the hybrid IR24×78-15. Our results indicate that the genetic distances from Xa-min(t) to ScBE05300 and BE061400 are 2.2 cM and 3.7 cM respectively on the same side. This study may facilitate the construction of the fine physical map of the Xa-min(t) gene. 相似文献