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Non-coding region and P.M gene sequence and vector construction of long-47 measles virus

A technology of measles virus and non-coding regions, applied in virus/bacteriophage, gene therapy, genetic engineering, etc., can solve the problem of protecting the intellectual property rights of MV vaccines, genome non-coding region sequences and P, M coding region sequences have not been elucidated and restricted Issues such as gene transfer and expression vectors

Inactive Publication Date: 2003-04-09
中国预防医学科学院病毒学研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Based on the many advantages of measles virus as a carrier, the long-47 live attenuated vaccine strain in my country was developed by traditional methods, and its safety and reliability have been tested and recognized for a long time, but its genome non-coding region sequence and P, M coding Region sequence has not been elucidated
On the one hand, this cannot protect the intellectual property rights of my country's MV vaccine at the genetic level, and on the other hand, it also limits the use of the vaccine virus to construct gene transfer and expression vectors

Method used

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  • Non-coding region and P.M gene sequence and vector construction of long-47 measles virus
  • Non-coding region and P.M gene sequence and vector construction of long-47 measles virus
  • Non-coding region and P.M gene sequence and vector construction of long-47 measles virus

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Embodiment 1

[0011] Example 1 The intergenic non-coding region sequence of the long-47 vaccine strain at the cDNA level can provide a site for inserting foreign genes into the entire virus genome for the construction of vaccine virus expression vectors, for the construction of genetic recombination multivalent vaccines and / Or gene therapy.

[0012] Measles virus is a member of the morbillivirus genus of the Paramyxoviridae family. It is a non-segmented single-stranded negative-strand RNA virus. The genome consists of six non-overlapping cistrons arranged in series: 3'-NPMFHL -5', encoding a total of 8 proteins: N, P / C / V, M, F, H, L, the two ends of the genome and the intergenic regions of each structure are non-coding regions. The long-47 measles vaccine strain has gene termination signals and gene initiation signal sequences bounded by conserved GAA trinucleotides in almost every non-coding region between structural genes (exception: CCC trinucleotides between H / L) Sequence), they control th...

Embodiment 2

[0013] Example 2 The 3'-end non-coding region and 5'-end non-coding region sequence of the measles virus long-47 vaccine strain genome can be used to construct a defective virus particle vector expressing foreign genes. The 3'-end non-coding region of the former genome contains the initiation signal of viral antigenome replication, the initiation signal of N protein mRNA transcription and the packaging signal that binds to the viral N protein, etc.; the latter's 5'-end non-coding region sequence contains The initiation signal of viral genome (Genome) replication, the termination signal of L protein mRNA transcription, and the packaging signal of binding virus N protein. Connect the 3'end of the 3'end non-coding region sequence cDNA to the downstream foreign gene start codon according to conventional cloning methods, and connect the 5'end of the 5'end non-coding region sequence cDNA according to conventional cloning methods The stop codon of the upstream exogenous gene is then tran...

Embodiment 3

[0014] Example 3 The coding region sequences of the P and M genes of the measles virus long-47 vaccine strain and the encoded amino acid sequences can be used to construct genetic recombinant antigens for serological diagnosis of measles virus infection or epidemiological investigation after vaccination.

[0015] According to the determined sequence of the P and M protein genes of the long-47 vaccine strain and the results of the amino acid sequence analysis results, the full gene sequence of the two or the gene containing the main epitope can be selected to construct a prokaryotic or eukaryotic expression vector , Express the corresponding M and P proteins in prokaryotic cells and eukaryotic cells, and after purification, they can be used for serological diagnosis of measles virus infection or epidemiological investigation after vaccination.

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Abstract

The present invention provides coding sequences and structures of all the non-coding regions and P and M coding regions of Longchun-47 measles virus, protein amino acid sequence and structure of P and M coding regions, the use of the non-coding region sequences in constituting the vaccine virus carrier and the constituting process of the carrier. These data and method may be used in recombining multiple effect vaccine or gene treatment via utilizing the vaccine strain virus to constitute expression vector and may be also used in molecular biological diagnosis and reagent development for measles virus infection.

Description

Invention field [0001] The present invention belongs to the field of virology and molecular biology, and relates to the discovery of all non-coding regions and P and M coding region sequences of a measles virus vaccine strain. Specifically, it clarifies a strain successfully developed in my country for measles control All non-coding region sequences of live attenuated vaccine strains, P, M coding region sequences and amino acid sequences of the encoded proteins and their application range. Background of the invention [0002] Measles Virus (MV) is a member of the morbillivirus genus of the Paramyxoviridae family. It is a non-segmented single-stranded negative-stranded RNA virus. It is also one of the viruses that cause highly infectious diseases in the world. Droplets through the respiratory tract, or contact infection. The incidence of measles in susceptible people is almost 100%. Most infected people can recover completely, but a few can cause severe respiratory or central nerv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/165A61K48/00C12N7/01C12N15/45
Inventor 李德新胡孔新王晓芳李川孟祥芝梁米芳
Owner 中国预防医学科学院病毒学研究所
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