Journal of molecular and applied genetics最新文献_第2页

Metaphase chromosome transfer of introduced selectable markers. 引入选择性标记的中期染色体转移。

Journal of molecular and applied genetics Pub Date : 1984-01-01

D L Nelson, J H Weis, M J Przyborski, R C Mulligan, J G Seidman, D E Housman

{"title":"Metaphase chromosome transfer of introduced selectable markers.","authors":"D L Nelson, J H Weis, M J Przyborski, R C Mulligan, J G Seidman, D E Housman","doi":"","DOIUrl":"","url":null,"abstract":"A general method for creating somatic cell hybrids which maintain chromosome segments by selection is described. To extend the technique of metaphase chromosome transfer to regions of the genome which do not carry a selectable marker, a retroviral vector has been employed to introduce the dominant selectable neo gene at many genomic locations in a population of murine cells. We have used such cells as donors in metaphase chromosome transfer experiments in which hamster and monkey cells were used as recipients. Cells acquiring a transferred chromosomal segment containing the neo gene were selected by growth in the presence of the drug G418. Hybridization to cloned interspersed repeat DNA sequences of the mouse was used to estimate the proportion of mouse DNA in each transferent. These experiments indicate that transferents produced in this manner contain 0.01 to 1.0% of the mouse genome. To analyze the organization of the DNA transferred to each recipient, we used Southern transfer hybridization of DNA from each transferent to a cloned mouse interspersed repeated DNA probe which did not cross-hybridize to hamster or monkey DNA. We found that each primary transferent gave a unique pattern of restriction fragments hybridizing to this probe. Secondary transferents from two independent primary transferents were compared by this technique. Each set of secondary transferents exhibited a hybridization pattern which resembled closely that of the primary transferent from which it was derived. However, a number of hybridizing DNA segments present in each primary transferent were absent in some of the secondary transferents. These results are most compatible with the view that an intact segment of the mouse chromosome surrounding the integration site of the retroviral vector can be transferred by the techniques used in this study. We believe this technique will be generally applicable to cells from many species, and will allow the study of chromosome regions previously refractory to analysis by chromosome transfer techniques.","PeriodicalId":77864,"journal":{"name":"Journal of molecular and applied genetics","volume":"2 6","pages":"563-77"},"PeriodicalIF":0.0,"publicationDate":"1984-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17165964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Affinity-purified antibody as a probe of RNA polymerase II subunit structure. 亲和纯化抗体作为RNA聚合酶II亚基结构的探针。

Journal of molecular and applied genetics Pub Date : 1984-01-01

A Robbins, W S Dynan, A Greenleaf, R Tjian

引用次数: 0

Analysis of two potential shuttle vectors containing herpes simplex virus defective DNA. 含单纯疱疹病毒缺陷DNA的两种潜在穿梭载体分析。

Journal of molecular and applied genetics Pub Date : 1984-01-01

S E Bear, A M Colberg-Poley, D L Court, B J Carter, L W Enquist

{"title":"Analysis of two potential shuttle vectors containing herpes simplex virus defective DNA.","authors":"S E Bear, A M Colberg-Poley, D L Court, B J Carter, L W Enquist","doi":"","DOIUrl":"","url":null,"abstract":"Two potential shuttle vectors which contained the identical herpes simplex virus type 1 (HSV-1) defective particle DNA (dDNA), but prokaryotic DNA of different origins, were examined for their stability when propagated in eukaryotic cells, and for their efficiency as shuttle vectors. Each chimeric molecule contained a 9.5 kilobase-pair (kb) EcoRI fragment (HSV12-7) representing a single unit of a class I HSV-1 dDNA. This dDNA was cloned into the bacteriophage lambda (lambda) vector lambda gtWES X lambda B' to create a 45.3 kb chimeric molecule (lambda gtWES::12-7), and into the plasmid vector pBR325, resulting in a 15.5 kb recombinant DNA molecule (pBR325::12-7). Each of these DNA molecules was transfected independently into African green monkey kidney cells which were then infected with wild-type HSV-1 helper virus. Both chimeric molecules were replicated and packaged into HSV-1 virions. However, regions of the lambda gtWES::12-7 chimeric DNA were rapidly deleted and rearranged, whereas the plasmid/HSV-1 DNA molecules were less rearranged. No intact lambda gtWES::12-7 DNA was recovered from HSV-1 virions as detected by infectivity of in vitro packaged DNA. However, pBR325::12-7 DNA isolated from HSV-1 virions was able to transform E. coli to ampicillin resistance. These results suggest additional considerations when designing single units of HSV-1 dDNA for use as vectors to accommodate large fragments of DNA.","PeriodicalId":77864,"journal":{"name":"Journal of molecular and applied genetics","volume":"2 5","pages":"471-84"},"PeriodicalIF":0.0,"publicationDate":"1984-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17157669","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Herbicide-resistant alfalfa cells: an example of gene amplification in plants. 抗除草剂苜蓿细胞:植物基因扩增的一个例子。

Journal of molecular and applied genetics Pub Date : 1984-01-01

G Donn, E Tischer, J A Smith, H M Goodman

引用次数: 0

The nifH and nifDK promoter regions from Rhizobium japonicum share structural homologies with each other and with nitrogen-regulated promoters from other organisms. 日本根瘤菌的nifH和nifDK启动子区域彼此具有结构同源性，并与其他生物的氮调控启动子具有结构同源性。

Journal of molecular and applied genetics Pub Date : 1984-01-01

T H Adams, B K Chelm

{"title":"The nifH and nifDK promoter regions from Rhizobium japonicum share structural homologies with each other and with nitrogen-regulated promoters from other organisms.","authors":"T H Adams, B K Chelm","doi":"","DOIUrl":"","url":null,"abstract":"In several species of Rhizobium the three genes which encode the nitrogenase complex are separated into two operons, nifH and nifDK. We have mapped the transcriptional promoter sites for these two operons from R. japonicum USDA strain 110 by S1 protection analyses using bacterial RNA isolated from soybean nodules. Transcription of the nifDK operon is initiated at a site located 46 nucleotides upstream of the proposed translation initiation codon. nifH transcription initiates predominantly at a site 152 nucleotides upstream of the proposed translation initiation codon. An additional minor start for nifH is found 35 nucleotides downstream of the major initiation site. The nucleotide sequences of these promoter sites are presented. Comparison of the major R. japonicum nif promoter sequences reveals a high degree of sequence homology with the conserved sequences clustered in the -11 to -15, -21 to -25, -27 to -31, and -38 to -41 regions. Conservation is also observed for the -11 to -15 and -21 to -25 regions with the R. meliloti nifHDK, a cowpea Rhizobium nifH, and even Klebsiella pneumoniae nif promoters. The -27 to -31 and -38 to -42 region homologies are not conserved with the other known nif promoters. These results are discussed in light of models for nif promoter activation.","PeriodicalId":77864,"journal":{"name":"Journal of molecular and applied genetics","volume":"2 4","pages":"392-405"},"PeriodicalIF":0.0,"publicationDate":"1984-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17644292","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Molecular characterization of a deletion mutation affecting the alpha'-subunit of beta-conglycinin of soybean. 影响大豆 beta 连环素 alpha'-亚基的缺失突变的分子特征。

Journal of molecular and applied genetics Pub Date : 1984-01-01

B F Ladin, J J Doyle, R N Beachy

{"title":"Molecular characterization of a deletion mutation affecting the alpha'-subunit of beta-conglycinin of soybean.","authors":"B F Ladin, J J Doyle, R N Beachy","doi":"","DOIUrl":"","url":null,"abstract":"A naturally occurring variety of soybean Glycine max cv. Keburi, which lacks the alpha'-subunits of the 7S seed storage protein (beta-conglycinin), was recently described by Kitamura and Kaizuma. Keburi beta-conglycinins contain a normal complement of alpha-, beta-, and gamma-subunits that accumulate in a temporally and spatially regulated manner comparable to that of the standard cultivar Provar. Poly(A)+ RNA isolated from mid-to-late maturation stage Keburi seeds was translated in vitro, yielding products equivalent to those of Provar poly(A)+ RNA except that Keburi mRNA did not produce the pre-alpha'-subunit polypeptide. The basis of the Keburi phenotype was determined by examining genomic DNA using Southern blot hybridization. Restriction fragments isolated from a cloned 11.5 kb EcoRI fragment of genomic DNA containing a normal alpha'-subunit gene (Gmg 17.1) were used as hybridization probes. Sequences far upstream of the alpha'-subunit gene were present in both Provar and Keburi cultivars. However, hybridization reactions with probes from within the gene demonstrated that a deletion had occurred in Keburi DNA beginning immediately 5' to the alpha'-subunit gene represented on this 11.5 kb fragment. The deletion extends through most of the coding sequences, producing the Keburi phenotype.","PeriodicalId":77864,"journal":{"name":"Journal of molecular and applied genetics","volume":"2 4","pages":"372-80"},"PeriodicalIF":0.0,"publicationDate":"1984-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17596182","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Nucleotide sequence and transcript mapping of the tmr gene of the pTiA6NC octopine Ti-plasmid: a bacterial gene involved in plant tumorigenesis. pTiA6NC章鱼碱ti质粒tmr基因的核苷酸序列和转录图谱:一个参与植物肿瘤发生的细菌基因。

Journal of molecular and applied genetics Pub Date : 1984-01-01

C Lichtenstein, H Klee, A Montoya, D Garfinkel, S Fuller, C Flores, E Nester, M Gordon

引用次数: 0

M13 bacteriophage vectors for the expression of foreign proteins in Escherichia coli: the rabies glycoprotein. M13噬菌体载体在大肠杆菌中表达外源蛋白:狂犬病糖蛋白。

Journal of molecular and applied genetics Pub Date : 1984-01-01

R F Lathe, M P Kieny, D Schmitt, P Curtis, J P Lecocq

引用次数: 0

Structure of a chromosomal Phaseolus vulgaris lectin gene and its transcript. 菜豆凝集素染色体基因的结构及其转录本。

Journal of molecular and applied genetics Pub Date : 1984-01-01

L M Hoffman