Bat-derived adeno-associated virus genome, amplification primers and their application, and amplification method

Abstract

The invention discloses a bat-derived adeno-associated virus genome, amplification primers and their application, and an amplification method. to the amplification primers, five pairs of primers are used herein as upstream primers and downstream primers; an adeno-associated virus RNA template extracted from a bat sample is subjected to RT-PCR (reverse transcription-polymerase chain reaction) amplification to obtain the amplification primers; the amplification primers are subjected to nucleotide sequencing; a complete sequence of the bat-derived adeno-associated virus genome is acquired by splicing and comparing. In allusion to characteristics of the bat-derived adeno-associated virus genome, short sequence fragments of the bat-derived adeno-associated virus genome are acquired by sequencing in reference to metagenomics in the lab; a fragmented amplification strategy is designed according to an open reading box included in the genome; the corresponding primers are designed according tothe conserved region; five pairs of primers are acquired by screening and are effective in amplifying genome sequencing of the bat-derived adeno-associated virus. The bat-derived adeno-associated virus genome amplification primers are widely applicable to the demand for detecting bat-derived adeno-associated viruses and to the corresponding field of scientific research.

Description

technical field [0001] The invention relates to a bat-derived adeno-associated virus genome, amplification primers and the application and amplification method thereof in the amplification of the bat-derived adeno-associated virus genome, belonging to the field of biotechnology. Background technique [0002] Adeno-associated viruses (AAVs) were discovered in 1965. During the preservation of adenoviruses, a small virus was found to be always together with adenoviruses, so it was named AAVs. Since it was initially found that AAVs could efficiently infect cells and replicate infected progeny only by co-infection with adenoviruses or herpesviruses, adeno-associated viruses were classified as a separate genus within the family Parvoviridae, the genus Dependovirus. In cultured normal cells, pure adeno-associated virus does not cause virulent infection, and can only produce effects when it is co-infected with other helper viruses. Adeno-associated virus is currently not considered...

AI Technical Summary

Problems solved by technology

Due to its good safety, wide host range, and low immunogenicity, adeno-associated virus is widely used in the construction of gene expression vectors and gene therapy, but its random integration may still cause potential dangers in clinical application.

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