No abstract available.
Clone Cells* ; Humans*

Compounds of polyketide origin possess a wealth of pharmacological effects, including antibacterial, antifungal, antiparasitic, anticancer and immunosuppressive activities. Many of these compounds and their semisynthetic derivatives are used today in the clinic. Most of the gene clusters encoding commercially important drugs have also been cloned and sequenced and their biosynthetic mechanisms studied in great detail. The area of biosynthetic engineering of the enzymes involved in polyketide biosynthesis has recently advanced and been transferred into the industrial arena. In this work, we introduce a computational system to provide the user with a wealth of information that can be utilized for biosynthetic engineering of enzymes involved in post-PKS tailoring steps. Post-PKS tailoring steps are necessary to add functional groups essential for the biological activity and are therefore important in polyketide biosynthesis.
Clone Cells ; Multigene Family

No abstract available.
Clone Cells* ; Humans* ; Lymphocytes*

No abstract available.
Animals* ; Clone Cells*

No abstract available.
Clone Cells* ; Microdissection*

No abstract available.
Clone Cells* ; Cloning, Organism* ; Nucleopolyhedrovirus*

No abstract available.
Clone Cells* ; Cloning, Organism* ; Thymidine*

White and brown strains of Flammulina velutipes were inter-crossed. All F1 showed light-brown fruitbody, suggesting that a gene for the brown fruitbody was incompletely dominant against the white one. And backcross experiment showed that more than two genes were involved in color determination. To isolate a molecular marker linked to fruitbody color, a set of primers was designed from a sequence of clones derived by a bulked segregant analysis. These markers showed a specific band which co-segregated with brown fruitbody forming strains.
Clone Cells ; Flammulina* ; Genes, vif

Hepatitis C virus (HCV) E2 protein is known to be one of putative envelope proteins. To develop a sensitive detection method for HCV infected tissues and cells, monoclonal antibodys (MAbs) to the E2 protein of HCV were prepared from mice immunized with recombinant baculovirus-expressing E2 protein (Bac-E2). Several hybridoma clones secreting various levels of MAb were isolated and isotypes of these MAb were determined. One clone (L.2.3.3) was used for ascites production and the E2-MAb was purified and characterized. The L.2.3.3 reacted well with both Bac-E2 and E. coli expressed glutathione-5-transferase-E2 (GST-E2) fusion proteins. Using HCV patient sera, E2 envelope protein was found to be localized in the cell membrane boundary both in CHO cells and insect cells which express HCV E2 protein. Similar result was obtained when same cells were treated with the MAb L.2.3.3. These results demonstrated that Bac-E2 protein is capable of eliciting high titer antibody production in mice.
Animals ; Antibody Formation ; Ascites ; Cell Membrane ; CHO Cells ; Clone Cells ; Cricetinae ; Hepacivirus* ; Hepatitis C* ; Hepatitis* ; Humans ; Hybridomas ; Insects ; Mice

BACKGROUND: Members belonging to the interferon-lambda (IFN-lambda) family exert protective action against viral infection; however, the mechanisms of their action have remained elusive. To study IFN-lambda biology, such as endocytosis of IFN-lambda, we produced monoclonal antibodies (Abs) against human IFN-lambda and examined their usefulness. METHODS: We purified recombinant human IFN-lambda1 expressed in Escherichia coli by using affinity columns. Then, we generated hybridoma cells by fusing myeloma cells with splenocytes from IFN-lambda1- immunized mice. For evaluating the neutralizing activity of the monoclonal Abs against IFN-lambda1, we performed RT-PCR for the MxA transcript. In order to study the binding activity of IFN-lambda and the monoclonal Ab complex on HepG2 cells, we labeled the monoclonal Ab with rhodamine and determined the fluorescence intensity. RESULTS: Four hybridoma clones secreting Abs specific to IFN-lambda1 were generated and designated as HL1, HL2, HL3, and HL4. All the Abs reacted with IFN-lambda1 in the denatured form as well as in the native form. Abs produced by HL1, HL3, and HL4 did not neutralize the induction of the MxA gene by IFN-lambda1. We also demonstrated the binding of the HL1 monoclonal anbitody and IFN-lambda complex on HepG2 cells. CONCLUSION: Monoclonal Abs against IFN-lambda1 were produced. These Abs can be used to study the cellular binding and internalization of IFN-lambda.
Animals ; Antibodies, Monoclonal ; Biology ; Clone Cells ; Endocytosis ; Escherichia coli ; Fluorescence ; Hep G2 Cells ; Humans ; Hybridomas ; Mice ; Rhodamines