摘要:
An expression vector for preparing a library of an antibody variable region, which can express comprising polypeptides containing the H-chain and/or L-chain variable regions of antibodies in the membrane-bound form on the surfaces of eukaryotic cells, which is containing the nucleotide sequence of AKL (formula 1) and/or the nucleotide sequence AKH (formula 2), and which is replicable in the cells: 5′-PL-SL-CL-ML-AL-3′ (formula 1)(AKL) 5′-PH-SH-CH-MH-AH-3′ (formula 2)(AKH) wherein PL and PH represent a promoter, respectively; SL and SH represent a nucleotide sequence coding for a signal peptide respectively; CL represents a nucleotide sequence coding for the L-chain constant region of an antibody; CH represents a nucleotide sequence coding for the H-chain constant region of an antibody or a nucleotide sequence coding for a polypeptide containing at least CH1 of the H-chain constant region of an antibody; AL and AH represent a polyadenylation signal, respectively; “5′-” and “-3′” refer to the 5′ side and the 3′ side, respectively, of the nucleotide sequence; and ML and MH represent a nucleotide sequence coding for the transmembrane domain, respectively, while either ML or MH may be a mere chemical bond when the expression vector contains the nucleotide sequence of AKL and AKH. Cloning site of R1L, R2L, R1H and R2H is present within or in the vicinity of SL, CL, SH and CH respectively, in order for the nucleotide sequences coding for the L-chain and H-chain variable regions to be readily inserted in between SL and CL, and SH and CH respectively.
摘要:
A method of separating and purifying nucleic acids from samples containing cells, such as blood and culture solutions. According to the method of the invention, a cell extract obtained by cell disruption is adsorbed by a filter made of a nonwoven fabric and the nucleic acid is eluted after washing the filter. Alkaline conditions of pH 12 or higher may be employed for elution of the nucleic acid, or the filter-adsorbed nucleic acid may be eluted by treatment with active oxygen or by using a surfactant. Nucleic acids separated and purified by the method of the invention can be used in nucleic acid amplification and nucleic acid sequence analysis techniques.
摘要:
Disclosed are a novel seven-pass transmembrane receptor protein found in immature dendritic cells and a DNA encoding the same. Further, disclosed are a replicable recombinant DNA which comprises a replicable expression vector and, operably inserted therein, the above-mentioned DNA; a cell of a microorganism or cell culture (transformant), which is transformed with the above-mentioned replicable recombinant DNA; a seven-pass transmembrane receptor protein which is produced on the cell surface of the above-mentioned transformant; a method for screening a ligand which binds to the above-mentioned seven-pass transmembrane receptor protein, and a method for screening a substance which inhibits the ligand from binding to the seven-pass transmembrane receptor protein; and an antibody which binds to the above-mentioned seven-pass transmembrane receptor protein. The present invention also discloses a method for the diagnosis of an inflammatory disease, such as rheumatism, which comprises determining the amount of the seven-pass transmembrane receptor protein expressed in human leukocytes.