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1.
Frederick J. Darfler 《In vitro cellular & developmental biology. Plant》1990,26(8):769-778
Summary A protein-free medium, termed ABC, has been developed which essentially eliminates the need for serum proteins. ABC supports
the long-term growth of murine hybridomas as well as other transformed cells of the immune system. The requirement of hybridoma
growth for transferrin has been met by substituting the soluble organo-iron compound, sodium nitroprusside. Substantial improvement
in the growth of hybridomas was afforded by the inclusion of 18 trace elements complexed to disodium ethylene diaminetetraacetate
(EDTA). The medium was further improved by the inclusion of components not found in Ham's F12 medium or by raising the concentrations
of existing low molecular weight components. Murine hybridomas can be cultured routinely in this protein-free medium in an
anchorage-independent manner with doubling times generally under 24 h. Visualized on electrophoretic gels, levels of monoclonal
antibody taken from those cultures often exceeded 80% of the total protein. The medium was also able to support the growth
of HuT 78 and H9 cells as well as certain other transformed cells of the immune system. In addition, normal human peripheral
blood lymphocytes, activated with phytohemagglutinin and cultured with 50 U/ml recombinant interleukin 2, could be grown for
2 wk with a 50-fold expansion over input cell number. 相似文献
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真空负压和常规ABC法显示HPV—1抗原的应用比较 总被引:3,自引:1,他引:2
应用真空负压 ABC 法显示 HPV-1抗原比常规 ABC 法效果好,敏感性高,时间短,整个过程只需一个多小时,阳性物明显突出,颗粒均匀,色泽鲜艳,呈黄棕色,背景清晰等优点。还可提高抗体的稀释度,降低成本。与微波技术相比,具有设备简单,易于操作,安全可靠,不受条件限制,经对26例尖锐湿疣的病例进行多层次稀释度的研究,取得了满意的结果,认为该法值得推广应用。 相似文献
4.
Patrick Diep Heping Leo Shen Julian A. Wiesner Nadia Mykytczuk Vladimiros Papangelakis Alexander F. Yakunin Radhakrishnan Mahadevan 《Engineering in Life Science》2023,23(7):2200133
Mine wastewater often contains dissolved metals at concentrations too low to be economically extracted by existing technologies, yet too high for environmental discharge. The most common treatment is chemical precipitation of the dissolved metals using limestone and subsequent disposal of the sludge in tailing impoundments. While it is a cost-effective solution to meet regulatory standards, it represents a lost opportunity. In this study, we engineered Escherichia coli to overexpress its native NikABCDE transporter and a heterologous metallothionein to capture nickel at concentrations in local effluent streams. We found the engineered strain had a 7-fold improvement in the bioaccumulation performance for nickel compared to controls, but also observed a drastic decrease in cell viability due to metabolic burden or inducer (IPTG) toxicity. Growth kinetic analysis revealed the IPTG concentrations used based on past studies lead to growth inhibition, thus delineating future avenues for optimization of the engineered strain and its growth conditions to perform in more complex environments. 相似文献
5.
The cross-immunoreactivity, topography, and fiber projections of the alpha MSH-immunoreactive specific neurons in the forebrain of the rat appear to be distinctly different from that of the neurons in the hypothalamic arcuate opiocortin system. The cell bodies, immunoreactive only to -MSH, have a specific pattern of distribution in the dorsal and lateral hypothalamic regions from the level of the retrochiasmatic region to the premammillary area of the posterior hypothalamus. Immunoreactive fibers of these cells appear to extend into regions of the cerebral cortex and hippocampus. An antomical relationship between the immunostained fibers and/or terminals of the arcuate opiocortin pool of neurons and the -MSH-immunoreactive perikarya is described utilizing the ABC (Avidin-Biotin-Peroxidase Complex) and ABC-GO (Glucose Oxidase) or glucose oxidase-antiglucose oxidase complex methods of immunocytochemistry in which two tissue antigens with contrasting colors are demonstrated in the same tissue section. 相似文献
6.
Karl Enquist Mawritz Fransson Carolina Boekel Inger Bengtsson Lisa Lang Sofia Johansson IngMarie Nilsson 《Journal of molecular biology》2009,387(5):1153-354
To what extent do corresponding transmembrane helices in related integral membrane proteins have different membrane-insertion characteristics? Here, we compare, side-by-side, the membrane insertion characteristics of the 12 transmembrane helices in the adenosine triphosphate-binding cassette (ABC) transporters, P-glycoprotein (P-gp) and the cystic fibrosis transmembrane conductance regulator (CFTR). Our results show that 10 of the 12 CFTR transmembrane segments can insert independently into the ER membrane. In contrast, only three of the P-gp transmembrane segments are independently stable in the membrane, while the majority depend on the presence of neighboring loops and/or transmembrane segments for efficient insertion. Membrane-insertion characteristics can thus vary widely between related proteins. 相似文献
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Sunanda Mallick Ashish Kumar Hiren Dodia Cyrus Alexander Dileep Vasudevan Tushar
Kant Beuria 《Bioscience reports》2021,41(1)
The peptidoglycan (PG) layer is an intricate and dynamic component of the bacterial cell wall, which requires a constant balance between its synthesis and hydrolysis. FtsEX complex present on the inner membrane is shown to transduce signals to induce PG hydrolysis. FtsE has sequence similarity with the nucleotide-binding domains (NBDs) of ABC transporters. The NBDs in most of the ABC transporters couple ATP hydrolysis to transport molecules inside or outside the cell. Also, this reaction cycle is driven by the dimerization of NBDs. Though extensive studies have been carried out on the Escherchia coli FtsEX complex, it remains elusive regarding how FtsEX complex helps in signal transduction or transportation of molecules. Also, very little is known about the biochemical properties and ATPase activities of FtsE. Because of its strong interaction with the membrane-bound protein FtsX, FtsE stays insoluble upon overexpression in E. coli, and thus, most studies on E. coli FtsE (FtsEEc) in the past have used refolded FtsE. Here in the present paper, for the first time, we report the soluble expression, purification, and biochemical characterization of FtsE from E. coli. The purified soluble FtsE exhibits high thermal stability, exhibits ATPase activity and has more than one ATP-binding site. We have also demonstrated a direct interaction between FtsE and the cytoplasmic loop of FtsX. Together, our findings suggest that during bacterial division, the ATPase cycle of FtsE and its interaction with the FtsX cytoplasmic loop may help to regulate the PG hydrolysis at the mid cell. 相似文献