Fluorescent clone screening vector and preparation and application thereof
A cloning vector and fluorescent screening technology, applied in the fields of molecular biology and microbiology, which can solve the problem of poor chromophore efficiency
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[0042] Construction and verification of embodiment 1 fluorescence screening cloning vector
[0043] The design idea and technical route of constructing the fluorescent screening cloning vector are as follows:
[0044] 1. Sequence design of enhanced green fluorescent protein gene. The green fluorescent protein gene in the vector pEGFP-N1 is EGFP, its detailed sequence is SEQ ID NO.2, and the corresponding amino acid sequence is SEQ ID NO.3, see the structure diagram image 3 . The crystal structure of GFP is a columnar structure composed of 11 β-sheets, and the GFP chromophore tyg is tightly packed in the β-box surrounded by β-sheets, which is protected from the surrounding environment. The present invention appropriately modifies the nucleic acid sequence without changing its protein structure, introduces a commonly used enzyme cutting site during the modification process, and modifies 4 amino acids at the same time, namely G20V, V29G, A37L, V55G ( Figure 4 ), the modifie...
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