genetic-transformation-mediated-agrobacterium-tumefaciens-solanum

Genetic Transformation Mediated by <em>Agrobacterium tumefaciens</em> in <em>Solanum nigrum</em>

Solanum nigrumis an orphan solanaceous relative of potato and tomato, and its fresh fruit is a nutritious and delicious berry. The whole plant of S. ...

Liaocheng University

A simplified Agrobacterium tumefaciens-mediated transformation protocol accelerates plant molecular breeding.

To clone DNA sequences quickly and precisely into plasmids is essential for molecular biology studies. Some cloning vectors have been developed for the cloning of PCR products, including blunt-end and T-A cloning. However, different plasmids are required for the cloning of PCR products with blunt ends and 3' A overhang ends. Here, a novel cloning vector, pYFRed, which is based on the pUC19 backbone, has emerged and can be applied in both blunt-end and T-A cloning. PCR products can be cloned into the pYFRed by a one-step digestion-ligation reaction in a tube. The endonuclease recognition sequences of I, 53kI, RV, I, and I for blunt-end cloning and I for T-A cloning were designed and added between the  promoter and the starting codon ATG of the  gene of pYFRed. The ligation efficiency was significantly higher because the restriction enzyme sites utilized were removed from the vector after being successfully constructed. The  gene was introduced into the pYFRed for the screening of the positive recombinants by the unaided eye without the need for additional reagents/equipment. pYFRed is easy to construct in an ordinary laboratory, which facilitates researchers to develop their cloning vector without purchasing commercial cloning vectors.

'Two in One' Cloning Vector Applied for Blunt-End and T-A Cloning with One-Step Digestion-Ligation and Screening of Positive Recombinants by Unaided Eyes.

Article	Year
A simplified Agrobacterium tumefaciens-mediated transformation protocol accelerates plant molecular breeding. Plant physiology\| PubMed ID: 39484976	2024
'Two in One' Cloning Vector Applied for Blunt-End and T-A Cloning with One-Step Digestion-Ligation and Screening of Positive Recombinants by Unaided Eyes. Current issues in molecular biology\| PubMed ID: 39852132	2024

Xingli Zhang

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