Construction and electrotransformation method of photosynthetic bacteria rhodopseudomonas palustris fluorescent GFP marker vector

Abstract

The invention discloses a vector recombination and construction method, a photosynthetic bacteria rhodopseudomonas palustris fluorescent GFP marker vector construction and electrotransformation method. The recombinant vector includes the photosynthetic bacteria rhodopseudomonas palustris fluorescent GFP marker vector including a PBBR1MCS-2 shuttle vector and a full-length fragment of a green fluorescent protein marker gene GFP, and is fused with a photosynthetic bacteria genome expression promoter PpckA gene and terminator TpckA gene, as well as a widely applied GFP gene, and a full-length fragment sequence of the GFP gene; the full-length fragment of the GFP gene is located between two ECORI and SACI restricted restriction enzyme sites of the PBBR1MCS-2 shuttle vector. The construction method includes the steps: genome DNA extraction, primer designing, PCR amplification, enzyme digestion, connection and other steps. The invention also includes obtained engineering bacteria containingthe recombinant vector, wherein the engineering bacteria are obtained by electrotransformation of the recombinant vector into competent cells. The engineering bacteria have the advantages of high stability, simple technology and environmental friendliness, and can be applied for study of a colonization behavior of photosynthetic bacteria in plants.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a recombinant vector and a construction method thereof, a fluorescent GFP-labeled photosynthetic bacterium Rhodopseudomonas palustris engineering bacteria, a preparation method and application thereof. Background technique [0002] Photosynthetic bacteria, as the earliest prokaryotic organisms that appeared on the earth and widely exist in nature, have primitive photosynthetic systems. Photosynthetic bacteria are Gram-negative bacteria, which can use natural inorganic and organic matter as nutrients to grow and reproduce under anaerobic light and aerobic dark conditions. In recent years, photosynthetic bacteria have been widely used in aquaculture, degradation of harmful substances, wastewater treatment, livestock and poultry production, and environmental protection because of their unique physiological functions and abundant bacteria. Many studies have shown that ph...

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Problems solved by technology

However, for photosynthetic bacteria, the col...

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