Reverse genetic manipulation method based on transcriptional regulatory sequence redistribution and application thereof

Abstract

The invention discloses a reverse genetic manipulation method based on transcriptional regulatory sequence redistribution and application thereof. The method comprises the steps that according to HuN4-F112 infectious clone full-length sequencing and identification of first-level core 6 base sequences of a transcriptional regulatory leader sequence (Leader TRS) and a downstream transcriptional regulatory sequence (body TRSs), a HuN4-F112 whole genome is designed to be six genome sequences containing Leader TRS and Body TRSs; primers for site-directed mutagenesis of Leader TRS and Body TRSs core 6 base sequences C5C6 are respectively designed according to six genome sequences including Leader TRS and Body TRSs, and genome segments of whole genome transcriptional regulation sequence overall mutation are obtained. A TRS circuit mutant PRRS attenuated vaccine strain can effectively resist a genome recombination phenomenon commonly occurring in a natural environment, and has important significance on effective prevention and control of PRRS clinically.

Description

technical field [0001] The invention belongs to the field of bioengineering, in particular, relates to a reverse genetic operation method based on the redistribution of transcription regulation sequences and its application, utilizes this method to construct virus mutant plasmids and genetic engineering vaccines, and more specifically relates to A mutant plasmid and genetic engineering vaccine capable of anti-recombination and effective immune protection against porcine reproductive and respiratory syndrome virus. Background technique [0002] Porcine Reproductive and Respiratory Syndrome, commonly known as "PRRS", clinically causes respiratory disorders in piglets, sow abortion and stillbirth. Huge economic loss. At present, live attenuated vaccine (MLV) immunity is common, and the infection rate of PRRSV is high. The disease is still the main disease in pig farms. The fattening pig farms in my country are generally positive, and there are only a handful of negative pig fa...

AI Technical Summary

Problems solved by technology

[0005] At present, in the clinical prevention and control of PRRSV, attenuated live vaccines are widely used, but the PRRSV genome mutation and recombination caused by the use of PRRSV have attracted extensive attention. The prevention and control of PRRSV has become more complicated and difficult, and the development is safer. Effective PRRSV attenuated live vaccine is imminent

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