Pseudorabies virus genome fosmid library, construction method and application in construction of mutants
A pseudorabies virus and a construction method technology, applied to the pseudorabies virus genome Fosmid library, construction and its application in the construction of mutants, can solve the problem of low efficiency of PRV mutants, inability to obtain recombinant viruses, affecting the efficiency of recombinant viruses, etc. problems, to achieve the effect of saving low efficiency, long construction cycle, and saving construction time
Active Publication Date: 2022-03-18
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
View PDF6 Cites 0 Cited by
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
However, the PRV genome is as high as 143kb, and it is very easy to break. Once the genome is broken, it loses infectivity and cannot obtain recombinant virus
In addition, the entire genome of PRV is connected with the BAC plasmid, and fragment deletions often occur during the electrotransformation process, which affects the efficiency of obtaining recombinant viruses (Lerma L, AL,Wagner S,Dinu M,Martín B,Tabarés E.Construction of recombinant pseudorabies viruses by using PRV BACsdeficient in IE180or pac sequences:Application of vBAC90D recombinant virus to production of PRV amplicons.Virus Res.2016,213:274,-282.) Therefore, the efficiency of obtaining PRV mutants by traditional homologous recombination methods is very low and takes a long time
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View moreImage
Smart Image Click on the blue labels to locate them in the text.
Smart ImageViewing Examples
Examples
Experimental program
Comparison scheme
Effect test
Embodiment Construction
[0040] The present invention will be further described below in conjunction with specific embodiments, and the advantages and characteristics of the present invention will become clearer along with the description. However, these embodiments are only exemplary and do not constitute any limitation to the scope of the present invention. It should be understood by those skilled in the art that the details and forms of the present invention can be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the protection scope of the present invention.
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More PUM
Login to View More Abstract
The invention discloses a pseudorabies virus (PRV) genome Fosmid library, a construction method and its application in the construction of PRV mutants. The present invention constructs the PRVTJ strain genome Fosmid library, selects 10 groups of 5-cosmid systems that can cover the complete PRV genome and have overlapping regions, screens out 5 groups of cosmids that can rescue the complete PRV, and identifies the best cosmid system. The cosmid combination was used to construct a recombinant PRV fused with VP26 to express EGFP. There was no difference between the reporter virus and the parental virus in titer and growth characteristics. The invention divides the huge PRV genome into several small fragments to construct a Fosmid library, which improves the stability of the library, makes the transformation and modification of the PRV gene simpler and more convenient, and shortens the cycle of building the library. The invention lays a foundation for rapid and accurate modification of the PRV genome, construction of recombinant vaccines and the like.
Description
technical field [0001] The present invention relates to a Fosmid library and a construction method thereof, in particular to a pseudorabies virus genome Fosmid library and a construction method thereof. The present invention further relates to the application of the pseudorabies virus genome Fosmid library in rapidly constructing pseudorabies virus mutants, which belongs to pseudorabies virus Genomic Fosmid library and its construction and application fields. Background technique [0002] Pseudorabies (Pseudorabies, PR) is a kind of pig, sheep, cattle and other livestock and wild animals caused by pseudorabies virus (Pseudorabies virus, PRV), with fever, itching (except pigs), encephalomyelitis, respiratory Acute infectious disease characterized by neurological and nervous system disorders (Mettenleiter, 1989). Pigs are the storage host and main source of infection of the disease, which can be transmitted horizontally through the digestive tract and respiratory tract, and c...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More Application Information
Patent Timeline
Login to View More Patent Type & Authority Patents(China)
IPC IPC(8): C40B50/06C40B40/06C12N7/01
CPCC12N7/00C40B40/06C40B50/06C07K14/43595C12N2710/16721
Inventor 仇华吉周末孙元
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI