Biomaterial Database

[Gene cloning and expression of mouse type I soluble interleukin 1 receptor in insect cells]. 1999

Z P Zhang, and Y Li

Institute of Medicinal Biotechnology, Chinese Academy of Medical Scienes & Peking Union Medical College, Beijing.

Interleukin 1(IL-1), which takes part in many physiological and pathological processes, is a kind of important cytokines. Their effects are exerted via specific IL-1 receptors (IL-1R) which are present on a wide variety of different cell types. Two types of IL-1R are identified as type I receptor (IL-1RI) and type II receptor (IL-1RII). Soluble forms of IL-1R can be produced from proteolytic cleavage of the extracellular portion of the two type receptors on cell surface. In this study, mouse type I soluble IL-1R (sIL-1RI) has been cloned and expressed in insect cells Sf9 with baculovirus as vector. Total RNA from NIH/3T3 cell was extracted with guanidinium thiocyanate followed by ultra-centrifugation in cesium chloride solution. The cDNA of sIL-1RI was amplified by RT-PCR technique. The cDNA was cloned into plasmid pAcGP67B. The recombinant plasmid pAI (pAcGP67B-sIL-1RI) was cotransfected with wild type AcNPV DNA into insect cells Sf9, because of genetic exchange occurred by homologous recombination in vivo, recombinant baculovirus rAcNPV was produced. Insect cells Sf9 were infected with the purified rAcNPV and sIL-1RI gene were expressed. SDS-PAGE analyses and Blockage assay of IL-1 beta biological activity demonstrated that the recombinant sIL-1RI had biological function and could be secreted into the medium.

UI	MeSH Term	Description	Entries
D011994	Recombinant Proteins	Proteins prepared by recombinant DNA technology.	Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D003001	Cloning, Molecular	The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.	Molecular Cloning
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D016475	3T3 Cells	Cell lines whose original growing procedure consisted being transferred (T) every 3 days and plated at 300,000 cells per plate (J Cell Biol 17:299-313, 1963). Lines have been developed using several different strains of mice. Tissues are usually fibroblasts derived from mouse embryos but other types and sources have been developed as well. The 3T3 lines are valuable in vitro host systems for oncogenic virus transformation studies, since 3T3 cells possess a high sensitivity to CONTACT INHIBITION.	3T3 Cell,Cell, 3T3,Cells, 3T3
D017472	Receptors, Interleukin-1	Cell surface receptors that are specific for INTERLEUKIN-1. Included under this heading are signaling receptors, non-signaling receptors and accessory proteins required for receptor signaling. Signaling from interleukin-1 receptors occurs via interaction with SIGNAL TRANSDUCING ADAPTOR PROTEINS such as MYELOID DIFFERENTIATION FACTOR 88.	IL-1 Receptor,IL-1 Receptors,IL1 Receptor,Interleukin-1 Receptor,Interleukin-1 Receptors,Receptor, Interleukin-1,Receptors, IL-1,IL1 Receptors,Interleukin 1 Receptor,IL 1 Receptor,IL 1 Receptors,Interleukin 1 Receptors,Receptor, IL-1,Receptor, IL1,Receptor, Interleukin 1,Receptors, IL 1,Receptors, IL1,Receptors, Interleukin 1
D051379	Mice	The common name for the genus Mus.	Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus
D018411	Spodoptera	A genus of owlet moths of the family Noctuidae. These insects are used in molecular biology studies during all stages of their life cycle.	Spodoptera frugiperda,Spodoptera frugiperdas,Spodopteras,frugiperdas, Spodoptera
D020133	Reverse Transcriptase Polymerase Chain Reaction	A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols.	Polymerase Chain Reaction, Reverse Transcriptase,Reverse Transcriptase PCR,PCR, Reverse Transcriptase,Transcriptase PCR, Reverse