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Recombinant spider poison protein, preparation method and expression vector thereof and medicament for treating (erection disturbance) ED

A kind of spider venom and protein technology, applied in the field of biochemistry

Inactive Publication Date: 2009-11-25
SICHUAN EMEISHAN PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to provide a recombinant spider venom, its preparation method, expression vector and medicine for treating ED, so as to solve the problem of spider venom in nature

Method used

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  • Recombinant spider poison protein, preparation method and expression vector thereof and medicament for treating (erection disturbance) ED
  • Recombinant spider poison protein, preparation method and expression vector thereof and medicament for treating (erection disturbance) ED
  • Recombinant spider poison protein, preparation method and expression vector thereof and medicament for treating (erection disturbance) ED

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] Embodiment 1. Use Escherichia coli to express and prepare the recombinant spider venom protein comprising PnTx2-6-α, which includes the following steps:

[0127] Artificially synthesized DNA fragments

[0128] 1 tcggcacgag atcagaatga aagttgcaat cctcttcctc tctattttgg tgcttgctgt

[0129] 61 tgcaagtgaa tccatgaag aatcccgtga tgattttgct gtagaagaat tggggagagc

[0130] 121 cacatgcgct ggccaagacc agccctgcaa agaaacttgc gactgctgtg gagagagg

[0131]181 agaatgtgtt tgcggaggac cttgcatttg caggcaaggc tacttttgga tagcatggta

[0132] 241 taaacttgct aactgtaaaa aatgatatgg atttatgtat accacgatat aataaatata

[0133] 301 ttgcatttga aaaaaaaaaaaaa;

[0134] Using the DNA fragment as a template, PCR amplifies the

[0135] tga aagttgcaat cctcttcctc tctattttgg tgcttgctgt

[0136] tgcaagtgaa tccattgaag aatcccgtga tgattttgct gtagaagaat tggggagagc

[0137] cacatgcgct ggccaagacc agccctgcaa agaaacttgc gactgctgtg gagagagagg

[0138] agaatgtgtt tgcggaggac cttgcatttg caggcaaggc tacttttgga tagcatggta ...

Embodiment 2

[0142] Example 2. Using Escherichia coli to express and prepare a recombinant spider venom protein comprising PnTx2-6-β, which includes the following steps:

[0143] Artificially synthesized DNA fragments

[0144] 1 gccacatgcg ctggccaaga ccagccctgc aaagaaactt gcgactgctg tggagagaga

[0145] 61 ggagaatgtg tttgcggagg accttgcatt tgcaggcaag gctacttttg gatagcatgg

[0146] 121 tataaacttg ctaactgtaa aaaatga

[0147] Using the DNA fragment as a template, PCR amplifies the

[0148] gc cacatgcgct ggccaagacc agccctgcaa agaaacttgc gactgctgtg gagagagagg

[0149] agaatgtgtt tgcggaggac cttgcatttg caggcaaggc tacttttgga tagcatggta

[0150] taaacttgct aactgtaaaaaatga

[0151] the nucleotide sequence.

[0152] The PCR product is recovered, and the sequence is inserted into the expression vector PSUMO-M at the multiple cloning site to obtain the expression vector of the recombinant PnTx2-6. Use the expression vector to transform DH5α competent cells, pick a single colony in the resistance p...

Embodiment 3

[0153] Example 3. Using yeast to express and prepare a recombinant spider venom protein comprising PnTx2-6-α, which includes the following steps:

[0154] Artificially synthesized DNA fragments

[0155] 1 tcggcacgag atcagaatga aagttgcaat cctcttcctc tctattttgg tgcttgctgt

[0156] 61 tgcaagtgaa tccatgaag aatcccgtga tgattttgct gtagaagaat tggggagagc

[0157] 121 cacatgcgct ggccaagacc agccctgcaa agaaacttgc gactgctgtg gagagagg

[0158] 181 agaatgtgtt tgcggaggac cttgcatttg caggcaaggc tacttttgga tagcatggta

[0159] 241 taaacttgct aactgtaaaa aatgatatgg atttatgtat accacgatat aataaatata

[0160] 301 ttgcatttga aaaaaaaaaaaaa

[0161] Using the DNA fragment as a template, PCR amplifies the

[0162] tga aagttgcaat cctcttcctc tctattttgg tgcttgctgt

[0163] tgcaagtgaa tccattgaag aatcccgtga tgattttgct gtagaagaat tggggagagc

[0164] cacatgcgct ggccaagacc agccctgcaa agaaacttgc gactgctgtg gagagagagg

[0165] agaatgtgtt tgcggaggac cttgcatttg caggcaaggc tacttttgga tagcatggta

[0166] taaac...

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Abstract

The invention provides a recombinant spider poison protein, a preparation method and an expression vector thereof and a medicament for treating (erection disturbance) ED so as to solve the problem of spider poison proteins in the nature. The recombinant spider poison protein consists of a polypeptide chain comprising the following sequence which consists of 48 amino acids: ATCAGQDQPC KETCDCCGER GECVCGGPCI CRQGYFWIAW YKLANCKK. The recombinant spider poison protein prepared by the invention can be used for treating the erection disturbance.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to a recombinant spider venom, its preparation method and expression vector, and a medicine for treating ED (male erectile dysfunction). Background technique [0002] Penile erection is a process of hemodynamic changes in the corpus cavernosum of the erectile organ under neuro-endocrine regulation. When sexually stimulated, non-adrenergic and non-cholinergic neurons synthesize and release nitric oxide (NO) under the catalysis of nitric oxide synthase (NOS); hexylcholine released from parasympathetic nerve endings activates the cavernous sinus of the penis And vascular endothelial cells nitric oxide synthase catalyzed to promote the synthesis and release of NO. NO diffuses into the smooth muscle cells of the corpus cavernosum and activates guanylate cyclase, which converts guanosine triphosphate into cyclic guanosine monophosphate (cGMP). The increase of intracellular cGMP concentration...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12C12N15/70C12N15/81C12N15/75C12N15/77C12N15/74C12N15/85C07K1/00A61K38/17A61P15/10
Inventor 胡浩高红
Owner SICHUAN EMEISHAN PHARMA
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