40 results about "Clone human" patented technology

Analogs of SRIF which are selective for SSTR1 in contrast to the other cloned SRIF receptors. These analogs are useful in determining the tissue and cellular expression of the receptor SSTR1 and its biological role in the endocrine, exocrine and nervous system, as well as in regulating tumor growth. SRIF analog peptides, such as des-AA^1,2,5[D-Trp⁸, N^αMeIAmp⁹, Tyr¹¹]-SRIF and counterparts incorporating Cbm at the N-terminus and/or N^αSer¹³, inhibit the binding of a universal SRIF radioligand to the cloned human receptor SSTR1, but they do not bind with significant affinity to human SSTR2, SSTR3, SSTR4 or SSTR5. By incorporating an iodinated tyrosine in position-2 or in position-11 in these SSTR1-selective SRIF analogs, a labeled compound useful in drug-screening methods is provided. The N-terminus accommodates bulky moieties without loss of selectivity, and a carbamoyl moiety or a conjugating agent that will accept a radioactive nuclide or will link to a cytotoxin may be present at the N-terminus.

Analogs of SRIF which are selective for SSTR4 in contrast to the other cloned SRIF receptors are useful in determining tissue and cellular expression of the receptor SSTR4 and its biological role in regulating tumor growth. SRIF analog peptides, such as des-AA^{1,2,4,5,12,13}[Ala⁷]-SRIF; des-AA^{1,2,4,5,12,13}[Aph⁷]-SRIF, des-AA^{1,2,4,5,12,13}[Aph⁷]Cbm-SRIF; des-AA^{1,2,4,5,12,13}[Tyr²,Ala⁷]-Cbm-SRIF, and des-AA^{1,2,4,5,12,13}[Tyr⁷,C^βMe-L-2Nal⁸]-SRIF, and counterparts incorporating D-Cys³ and/or D-Trp⁸ and/or Ala¹¹, bind with high affinity to the cloned human receptor SSTR4 and activate the receptor, but they do not bind with significant affinity to human SSTR1, SSTR2, SSTR3 or SSTR5. By incorporating an iodinated tyrosine in position-2 in these SSTR4-selective SRIF analogs, a labeled compound useful in drug-screening methods is provided. Alternatively, for use in therapy, cytotoxins or highly radioactive elements can be N-terminally coupled or complexed thereto.

The invention discloses a gene promoter of ginseng PgPDR3 responded by methyl jasmonate and an application thereof. The sequences of the gene promoter of the ginseng PgPDR3 are shown in SEQ ID NO.1, or are the DNA sequences of which the homology with SEQ ID NO.1 is over 99%. According to the invention, a Genomewalker technology is adopted to clone, later the sequences of the gene promoter of the PDR transport protein is obtained, and then the expression vector Pcambia1301-ProPDR3::GUS of the promoter is built; if the promoter in the transgenic tobacco drives the GUS to express, and can be controlled by ginsenoside, salicylic acid, gibberellins and abscisic acid, the promoter participates in the transport and accumulation of the ginsenoside, which provides a basis for cloning the gene of key control recording factor in the ginsenoside transport signal pathway and controlling the transport and accumulation of the terpenoid, such as ginsenoside; and fine plants containing rich ginsenoside can be obtained by using a method for controlling or improving the ginseng and other plants via the promoter or the derivative thereof.

SRIF peptide antagonists, which are selective for SSTR2 in contrast to the other cloned SRIF receptors and which bind with high affinity to the cloned human receptor SSTR2 but do not activate the receptor, have many useful functions. Because they do not bind with significant affinity to SSTR1, SSTR3, SSTR4 or SSTR5, their administration avoids potential undesirable side effects. Because they block the receptor function, they can be used therapeutically to block certain physiological effects which SSTR2 mediates. By incorporating radioiodine or the like in these SSTR2-selective SRIF antagonists, a labeled compound useful in drug-screening methods is provided. Alternatively, for use in therapy, highly radioactive moieties can be N-terminally coupled, complexed or chelated thereto.

A recombinant adeno-associated virus expressing antisense human CYP2J2 gene and preparation method thereof are provided, the recombinant adeno-associated virus is prepared through cloning human CYP2J2 cDNA (1509 bp, which encode the protein containing 503 amino acids) from human leucocyte DNA by PCR, cotransfecting three plasmids by the calcium phosphate precipitation technique to pack and prepare the recombinant adeno-associated virus containing the antisense target gene, and purifying. The recombinant adeno-associated virus obtained from the above method can be transfected into different kinds of human tumor cell lines, inhibiting the proliferation and migration of tumor cells, promoting the apoptosis of tumor cells and suppressing the growth and metastases of tumor. Therefore, it can prove that the selective inhibitor of CYP2J2 and the recombinant adeno-associated virus expressing antisense human CYP2J2 gene are the potential medicine for treating tumor.

The invention discloses a Pichia pastoris transformant capable of expressing human recombinant interleukin-3 (rhIL-3) with high efficiency and a purification method thereof. In the method, the eukaryotic host is pichia pastorisX-33. The purification method comprises the following steps: cloning a human IL-3 gene; establishing a eukaryotic expression vector, and transforming the eukaryotic expression vector into the eukaryotic yeast host; obtaining a yeast transformant for high-level secretory expression by screening, wherein the IL-3 expressed by the yeasts are available in a glycosylated mode and a non-glycosylated module; and performing amplified culture by using a shake flask, and subjecting the supernate of the culture solution to dialysis, nickel affinity purification and further purification by diethylaminoethanol (DEAE) anion column. The purified product is subjected to mass spectrometric identification and analysis, and the result of the mass spectometric identification and analysis indicates that the expressed IL-3 is modified by different glycosyls and that the IL-3 has an his*6 tag and a C-MYC tag and is easy for purification and detection of expression product. In the invention, different from the conventional method using a prokaryotic host to express the rhIL-3, the method for expressing a large amount of rhIL-3 by using a Pichia pastoris expression system is adopted for the first time, quick purification is realized by using a His-tag protein, the purified rhIL-3 is high-activity rhIL-3 protein which is glycosylated to different extents and of which the molecular weight is 19kDa and 22kDa. The method ensures that the high-activity rhIL-3 recombinant protein is obtained quickly.

The invention belongs to the technique field of producing multi-peptide drugs by gene engineering. It clones human EGF gene into the silkworm bacilliform virus (Bombyx mori Nuclear polyhedrosis Virus) transferring carrier pBacPAK8 and makes them carry through homologous reformation in the silkworm cells to obtain the reformed virus BmBacEGF with human-coat growing gene. Use the reformed virus to to inocualte the silkworm grub and chrysalis by acupuncture in order to make the human-coat growing gene efficient expressed. Then make the grub and chrysalis into oral drugs which has remarkable action on the digestible ulcer by examination on animal.

A cloned of a human brain lysophospholipid-specific lysophospholipase enzyme molecule, its potential use for treatment of a host of diseases and method of inactivation are disclosed. Also disclosed are its distribution in tissue sand detailed kinetic analysis. hLysoPLA has a single substrate binding site and a surface recognition site. In contrast to many nonspecific lipolytic enzymes that exhibit lysophospholipase activity, hLysoPLA hydrolyzes only lysophospholipids and has no other significant enzymatic activity.

The invention discloses a disease gene animal model of primary open angle glaucoma and a construction method thereof. The method comprises the following steps: (1) cloning human an OPTN (optineurin, E50K) mutant gene to an expression vector with a retinal specific promoter c-kit to construct a recombinant mammal expression vector; and (2) introducing the recombinant mammal expression vector into the bodies of mice and screening and identifying to obtain transgenic mice. In consideration of the difference between the mouse OPTN (E50K) and the human gene, the human OPTN (E50K) is introduced into the retinal specific promoter so that the human OPTN (E50K) gene is specifically expressed in the retina of the mouse. The model of primary open angle glaucoma due to the mutation of OPTN (E50K) is constructed, and the laboratory animals are provided for the systemic researches on pathogenic mechanism, development trend and disease prognosis and treatment of primary open angle glaucoma (POAG) due to the mutation of OPTN (E50K) in human beings.

This invention discloses a method for preparing trans-gene cultivated silkworm with human granulocyte, namely macrophage colony stimulating factor, characterized by the following steps: a, through gene engineering operation, cloning human granulocyte under the control of cultivated silkworm's karyotype polyhedrosis virogene promoter into a carrier with reporting gene based on piggyBAC factor; b, constructing and recombining trans-gene carrier; c, at the presence of subsidiary particles, attaining silkworm with reporting gene by spermium conducting method, and testing out human granulocyte, breeding to silkworms.

The invention relates to an optimized porcine circovivus type 2 recombinant adenovirus construction method. The construction of recombinant adenovirus is completed by cloning Human cytomegalovirus first intron (Intron A) and woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) into an adenovirus shuttle vector. The construction method provided by the invention has the advantages that a Cap expression quantity is improved, so that adenovirus usage dose and adenovirus protein autoimmune response can be relieved and the preparation efficiency of the porcine circovivus type 2 recombinant adenovirus can be improved.

The invention relates to protein, in particular to human recombinant amelogenin and a preparation method thereof. The human recombinant amelogenin is prepared by the following method: a, cloning human amelogenin mature peptide gene hAm coded by the X chromosome; b, constructing recombinant prokaryotic expression plasmid PGEX4T1-hAm containing the human amelogenin mature peptide gene; c, transfecting the recombinant prokaryotic expression plasmid PGEX4T1-hAm obtained from the step b into an expression strain E.coli.Rosetta; d, inducing the cloning expression strain obtained from the step c to express fusion protein GST-hAm; and e, purifying the fusion protein by GST affinity chromatography. The invention firstly obtains the encoding sequence of human amelogenin mature peptide from human deciduous dental germ through cloning, and successfully expresses the fusion protein in bacillus through the prokaryotic expression plasmid; and the artificially synthesized recombinant amelogenin can be obtained through the purification by the GST affinity chromatography.

A transgenic vaccine for lung cancer is prepared through targeting the cloned human IFN-gamma whole length CDMA to the cell strain A548 of human pulmonary adenocarcinoma by retroviral rector PLXSN, resistance screen to obatain cellstrain hu gamma-IFN-A549, and conventional culture to obtain the vaccine. It can prevent recurrence and transfer of tumor.

The invention discloses a method for obtaining human alpha-fetoprotein, which comprises cloning the human alpha-fetoprotein gene into insect cell expression vectors pFastBac 1, pFastBac Dual or pFastBac1‑Gus, and transforming DH10Bac to obtain human alpha-fetoprotein-containing Bacmid gene, Bacmid transfected insect cells 2 to 4 days after the secretion of human alpha-fetoprotein into the culture medium, human alpha-fetoprotein secreted by nickel column and gel chromatographic column after purification can be active and extracted from human embryonic blood of alpha-fetoprotein. The invention utilizes the eukaryotic expression of Sf9 and Sf21 insect cells to obtain human alpha-fetoprotein with better activity.

The invention discloses a human platelet-derived growth factor (PDGF) A homologous dimers and a high-efficiency production method thereof. The method mainly comprises the following steps: cloning a human platelet-derived growth factor A spliceosome 2 gene; constructing an eukaryotic expression vector and transferring the eukaryotic expression vector into an eukaryotic yeast host; screening a yeast transformant for high level secretion expression, wherein the PDGF-As expressed by the yeast are in a glycosylated form and a non-glycosylated form; and performing amplified culture in a shake flask, dialyzing supernate of culture solution, purifying the supernate efficiency by using a chromatography (CM) cation exchange column and G-75 molecular sieve, and obtaining the novel modified protein of the PDGF-AA dimers, of which one PDGF-A monomer is glycosylated and of which the other PDGF-A monomer is not glycosylated, by using a CM cation exchange process and optimizing the pH value of dislysate, wherein the separated and purified new glycosylation modified protein of the PDGF-AA dimers has high uniformity.

The invention discloses human marrow-interstitial stem cell modified by TPO gene, its preparing process and use. The human marrow-interstitial stem cell is obtained by recombination gland relevant virus meso-guide TPO gene decoration. The preparation method comprises the following steps consequently cloning human tyre liver TPO gene, constructing carrier plasmid Paav-tpo-ires-hrGFP, preparing Raav-TPO viral vectors, culturing human marrow-interstitial stem cell, transfecting Raav-TPO on human marrow-interstitial stem cell. The human marrow-interstitial stem cell modified by TPO gene or human marrow-interstitial stem cell exudate modified by TPO gene can be used for preparing medicine of urging blood platelet-generating. The MSCs modified by TPO gene generates internal source TPO through cell secreting for treating blood platelet reduction so the side-effect generated from injection human recombination TPO is not generated.

The present invention discloses the expression of human SP-Al in Pichia pastoris. Said method is characterized by that it clones human SP-Al gene into expression vector, and utilizes electric shock transformation technique to transfer in into pichia pastoris, and screens out multicopy transformant from transformed strain, and utilizes high-expression methanol oxidase 1 (AOXI) to gene promotor, and uses methanol as unique carbon source to make induction expression of recombinant P. pastoris, in the described inductive expression the concentration of methanol is 0.1-1.0% (g/ml). Said ivnention can produce lots of high-activity human SP-Al, its maximum expression amount can be up to 150 mg/L,l and the expressed object protein can possess opsonic action for making macrophage to kill pathogen.