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Related Concept Videos

DNA-only Transposons02:57

DNA-only Transposons

DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
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Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX
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Published on: June 27, 2017

A versatile zero background T-vector system for gene cloning and functional genomics.

Songbiao Chen1, Pattavipha Songkumarn, Jianli Liu

  • 1Department of Plant Pathology, The Ohio State University, Columbus, Ohio 43210, USA.

Plant Physiology
|May 1, 2009
PubMed
Summary

A novel zero background TA cloning system offers a simple, cost-effective method for direct DNA fragment cloning. This high-throughput platform complements existing technologies for gene cloning and functional genomics.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Plant Science

Background:

  • High-throughput gene cloning and functional analysis are crucial due to advances in genomic sequencing.
  • Existing methods like Gateway cloning are effective but involve time-consuming and expensive two-step processes.

Purpose of the Study:

  • To develop a zero background TA cloning system for simple, high-efficiency direct cloning of PCR-amplified DNA fragments.
  • To create a cost-efficient and versatile platform for gene cloning and functional genomics applications.

Main Methods:

  • Utilized the restriction enzyme XcmI to generate T-overhangs on vectors.
  • Incorporated the negative selection marker gene ccdB to minimize vector self-ligation.
  • Developed specialized vectors for transient and stable transformation in plants.

Main Results:

  • Achieved zero background cloning with minimal self-ligation of DNA fragments.
  • Demonstrated the system's feasibility for various plant functional analyses, including gene expression, silencing, and protein localization.
  • Successfully created a versatile set of vectors for diverse applications.

Conclusions:

  • The developed zero background TA cloning system provides a general, cost-efficient, and high-throughput alternative to existing cloning methods.
  • This platform complements the Gateway cloning system, enhancing gene cloning and functional genomics research.
  • The system's adaptability allows for the development of specialized vectors for various organisms.