Strategies for Mutagenesis and Gene Cloning Using Transposon Tagging and T-DNA Insertional Mutagenesis

1992 ◽  
Vol 43 (1) ◽  
pp. 49-78 ◽  
Author(s):  
V Walbot
Keyword(s):  
Gene Cloning ◽  
Insertional Mutagenesis ◽  
Transposon Tagging ◽  
Dna Insertional Mutagenesis

scholarly journals Embryonic Lethals and T-DNA Insertional Mutagenesis in Arabidopsis

1991 ◽  
Vol 3 (2) ◽  
pp. 149 ◽  
Author(s):  
Deena Errampalli ◽  
David Patton ◽  
Linda Castle ◽  
Leigh Mickelson ◽  
Karl Hansen ◽  
...  
Keyword(s):  
Insertional Mutagenesis ◽  
Dna Insertional Mutagenesis

Insertional mutagenesis in Bacillus subtilis: mechanism and use in gene cloning

Gene ◽  
1982 ◽  
Vol 19 (3) ◽  
pp. 277-284 ◽  
Author(s):  
B. Niaudet ◽  
A. Goze ◽  
S.D. Ehrlich
Keyword(s):  
Bacillus Subtilis ◽  
Gene Cloning ◽  
Insertional Mutagenesis

scholarly journals Chlamydomonas WDR92 in association with R2TP-like complex and multiple DNAAFs to regulate ciliary dynein preassembly

2018 ◽  
Vol 11 (9) ◽  
pp. 770-780 ◽  
Author(s):  
Guang Liu ◽  
Limei Wang ◽  
Junmin Pan
Keyword(s):  
Mass Spectrometry ◽  
Insertional Mutagenesis ◽  
The Other ◽  
Cytoplasmic Protein ◽  
Flagellar Motility ◽  
Heavy Chains ◽  
The Third ◽  
Axonemal Dynein ◽  
Dynein Arms ◽  
Dna Insertional Mutagenesis

Abstract The motility of cilia or eukaryotic flagella is powered by the axonemal dyneins, which are preassembled in the cytoplasm by proteins termed dynein arm assembly factors (DNAAFs) before being transported to and assembled on the ciliary axoneme. Here, we characterize the function of WDR92 in Chlamydomonas. Loss of WDR92, a cytoplasmic protein, in a mutant wdr92 generated by DNA insertional mutagenesis resulted in aflagellate cells or cells with stumpy or short flagella, disappearance of axonemal dynein arms, and diminishment of dynein arm heavy chains in the cytoplasm, suggesting that WDR92 is a DNAAF. Immunoprecipitation of WDR92 followed by mass spectrometry identified inner dynein arm heavy chains and multiple DNAAFs including RuvBL1, RPAP3, MOT48, ODA7, and DYX1C. The PIH1 domain-containing protein MOT48 formed a R2TP-like complex with RuvBL1/2 and RPAP3, while PF13, another PIH1 domain-containing protein with function in dynein preassembly, did not. Interestingly, the third PIH1 domain-containing protein TWI1 was not related to flagellar motility. WDR92 physically interacted with the R2TP-like complex and the other identified DNNAFs. Our data suggest that WDR92 functions in association with the HSP90 co-chaperone R2TP-like complex as well as linking other DNAAFs in dynein preassembly.

T-DNA insertional mutagenesis in Arabidopsis

1992 ◽  
Vol 20 (5) ◽  
pp. 963-976 ◽  
Author(s):  
Csaba Koncz ◽  
Kinga N�meth ◽  
George P. R�dei ◽  
Jeff Schell
Keyword(s):  
Insertional Mutagenesis ◽  
Dna Insertional Mutagenesis

Genetic Dissection of T-DNA Insertional Mutants Reveals Uncoupling of Dikaryotic Filamentation and Virulence in Sugarcane Smut Fungus

2021 ◽  
Author(s):  
Shan Lu ◽  
Feng Guo ◽  
Zhiqiang Wang ◽  
Xiaorui Shen ◽  
Yizhen Deng ◽  
...  
Keyword(s):  
Regulatory Networks ◽  
Insertional Mutagenesis ◽  
Genetic Regulation ◽  
Genetic Dissection ◽  
Opposite Mating Type ◽  
Basidiomycetous Fungus ◽  
Sporisorium Scitamineum ◽  
Type Strains ◽  
Dna Insertional Mutagenesis ◽  
Teliospore Development

The biotrophic basidiomycetous fungus Sporisorium scitamineum causing smut disease in sugarcane is characterized by a life-cycle composed of a yeast-like nonpathogenic haploid basidiosporial stage outside the plant and filamentous pathogenic dikaryotic hyphae within the plant. Under field conditions, dikaryotic hyphae are formed after mating of two opposite mating-type strains. However, the mechanisms underlying genetic regulation of filamentation and its association with pathogenicity and development of teliospores are currently unclear. This study has focused on the characterization and genetic dissection of haploid filamentous mutants derived from T-DNA insertional mutagenesis. Our results support the existence of at least three genotypes among the six haploid filamentous mutants that differentially contribute to virulence and development of the whip and teliospore, providing a novel foundation for further investigation of the regulatory networks associated with pathogenicity and teliospore development in S. scitamineum.

scholarly journals Transposon tagging using Ty elements in yeast.

Genetics ◽  
1988 ◽  
Vol 120 (1) ◽  
pp. 95-108
Author(s):  
D J Garfinkel ◽  
M F Mastrangelo ◽  
N J Sanders ◽  
B K Shafer ◽  
J N Strathern
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Cloning ◽  
Molecular Analysis ◽  
Target Gene ◽  
Transposon Mutagenesis ◽  
Transposon Tagging ◽  
Chromosomal Mapping ◽  
Ty Elements ◽  
His3 Gene

Abstract We have used the ability to induce high levels of Ty transposition to develop a method for transposon mutagenesis in Saccharomyces cerevisiae. To facilitate genetic and molecular analysis, we have constructed GAL1-promoted TyH3 or Ty917 elements that contain unique cloning sites, and marked these elements with selectable genes. These genes include the yeast HIS3 gene, and the plasmid PiAN7 containing the Tn903 NEO gene. The marked Ty elements retain their ability to transpose, to mutate the LYS2, LYS5, or STE2 genes, and to activate the promoterless his3 delta 4 target gene. Ty elements containing selectable genes are also useful in strain construction, in chromosomal mapping, and in gene cloning strategies.

scholarly journals Cloning of flagellar genes in Chlamydomonas reinhardtii by DNA insertional mutagenesis.

Genetics ◽  
1993 ◽  
Vol 135 (2) ◽  
pp. 375-384 ◽  
Author(s):  
L W Tam ◽  
P A Lefebvre
Keyword(s):  
Insertional Mutagenesis ◽  
Genetic Model ◽  
Nitrate Reductase Gene ◽  
New Approach ◽  
Cellular Processes ◽  
Reductase Gene ◽  
Wide Range ◽  
Mutant Strains ◽  
Mutant Cells ◽  
Dna Insertional Mutagenesis

Abstract Chlamydomonas is a popular genetic model system for studying many cellular processes. In this report, we describe a new approach to isolate Chlamydomonas genes using the cloned nitrate reductase gene (NIT1) as an insertional mutagen. A linearized plasmid containing the NIT1 gene was introduced into nit1 mutant cells by glass-bead transformation. Of 3000 Nit+ transformants examined, 74 showed motility defects of a wide range of phenotypes, suggesting that DNA transformation is an effective method for mutagenizing cells. For 13 of 15 such motility mutants backcrossed to nit- mutant strains, the motility phenotype cosegregated with the Nit+ phenotype, indicating that the motility defects of these 13 mutants may be caused by integration of the plasmid. Further genetic analysis indicated that three of these mutants contained alleles of previously identified loci: mbo2 (move backward only), pf13 (paralyzed flagella) and vfl1 (variable flagellar number). Three other abnormal-flagellar-number mutants did not map to any previously described loci at which mutations produce similar phenotypes. Genomic sequences flanking the integrated plasmid in the mbo2 and vfl1 mutants were isolated and used as probes to obtain wild-type genomic clones, which complemented the motility defects upon transformation into cells. Our results demonstrate the potential of this new approach for cloning genes identified by mutation in Chlamydomonas.

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