Transforming animal and plant genes to make them glow in the dark is a complex process that involves knowledge and technology in many fields. The main steps include selecting suitable luminescent genes, constructing vectors, introducing target animal and plant cells, screening and identification. The following is a detailed introduction.： ### Choose the right luminescent gene -**Fluorescent protein gene**: There are many fluorescent protein genes in nature, such as green fluorescent protein (GFP), red fluorescent protein (RFP), etc. The proteins encoded by these genes can absorb specific wavelengths of light and emit different colors of fluorescence. -**Selection basis**: Select the appropriate fluorescent protein gene according to actual needs. For example, if you want the plant to emit green light, you can choose the GFP gene; if you need red light, choose the RFP gene. At the same time, factors such as gene expression efficiency, stability, and influence on the original physiological functions of animals and plants need to be considered. ### Construct gene expression vector -**Vector selection**: Choosing the right vector is the key to gene introduction. Commonly used vectors are plasmids, viral vectors, etc. Plasmid is a small cyclic double-stranded DNA molecule with multiple restriction enzyme cleavage sites and screening marker genes to facilitate the insertion and screening of exogenous genes. Viral vectors have the ability to efficiently infect cells and can effectively introduce exogenous genes into animal and plant cells. -**Component assembly**: The selected luminescent gene, promoter, terminator and other regulatory elements are connected to the vector to construct a complete gene expression vector. Promoter is a key regulatory element of gene expression, which can initiate the transcription process of genes. The terminator is located at the end of the gene, which can terminate the transcription process and ensure the accuracy of gene expression. ### Import gene expression vectors into target animal and plant cells -**Plant cells** -**Agrobacterium-mediated method**: This is one of the most commonly used methods in plant genetic engineering. The constructed gene expression vector is introduced into Agrobacterium, and then agrobacterium is used to infect plant cells and integrate exogenous genes into the plant genome. -**Gene marksmanship**: Particles wrapped in gene expression vectors are directly injected into plant cells through high-speed micro-bullets to achieve the introduction of exogenous genes. -**Animal cells** -**Microinjection method**: Using a micromanipulator, the gene expression vector is directly injected into the prokaryote of an animal fertilized egg to integrate exogenous genes into the animal genome. This method is suitable for all kinds of animal cells, but the operation technology requirements are high, and professional equipment and skilled operators are required. -**Virus-mediated method**: The use of viral vectors to infect animal cells and introduce exogenous genes into cells. Commonly used viral vectors are retroviral vectors, adenoviral vectors, etc. This method has a high infection efficiency, but there may be safety issues with viral vectors. ### Screening and identification of genetically modified animals and plants -**Screening markers**: Gene expression vectors usually have screening marker genes, such as antibiotics resistance genes. By adding corresponding antibiotics to the medium, cells or plants containing exogenous genes can be screened. -**Molecular identification**: Molecular biology techniques such as PCR, Southern hybridization, and fluorescence quantitative PCR are used to identify whether exogenous genes are integrated into the genomes of animals and plants, the sites of integration, and their expression. Only genetically modified animals and plants identified as positive can be used for subsequent research and application. Transforming the genes of animals and plants to make them glow in the dark is a challenging task that requires multi-disciplinary knowledge and technical support. When genetic modification is carried out, relevant laws, regulations and ethical guidelines must be strictly observed to ensure biological safety and ecological balance.