Abstract：
:The pLKO.1-TRC plasmid has been a popular and widely used vector due to its simple handling and stability. The huge RNAi database, a TRC library, has been established based on this vector. However, this plasmid only has a puromycin-resisted gene for selecting, which limits its application in microscopy and fluorescence-activated cell sorting (FACS). In the present work, PCR, restriction endonuclease digestion and molecular cloning techniques were used to insert the gene decoding green fluorescent protein (GFP) without changing the structure of original plasmid to extend its application. To demonstrate the function of new plasmid, we constructed shNC and shGAPDH plasmids based on newly constructed pLKO-TurboGFP-TRC (pLKOG) and original pLKO plasmids, and then packaged lentivirus particles by 293T cells. The supernatant containing lentiviral particles was collected and then incubated with RAW264.7 cells for infection. After selection for 7 days by using puromycin, the cells were harvested. RT-qPCR and Western blotting were used to detect the target gene expression. FACS was used to detect the green fluorescent of cells. Our results showed that the newly constructed pLKOG plasmid, as a lentiviral vector carrying shRNA, could knock down the target gene expression efficiently and express TurboGFP protein efficiently in the host cells. We conclude that the new plasmid is a convenient vector for selecting positive cells with shRNA by using fluorescent microscope and FACS.
journal_name
Appl Biochem Biotechnoljournal_title
Applied biochemistry and biotechnologyauthors
Xu J,Zhu W,Guo Y,Jiang C,Hussain N,Meng L,Lu Sdoi
10.1007/s12010-016-2242-1subject
Has Abstractpub_date
2017-02-01 00:00:00pages
699-709issue
2eissn
0273-2289issn
1559-0291pii
10.1007/s12010-016-2242-1journal_volume
181pub_type
杂志文章abstract：:Sweet sorghum (Sorghum sp.) has high biomass yield. Hydrolysis of lignocellulosic sweet sorghum bagasse (SSB) to fermentable sugar could be useful for manufacture of biofuel or other fermentation products. Pretreatment of lignocellulosic biomass to degrade lignin before enzymatic hydrolysis is a key step. Fungal pretr...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-017-2439-y
更新日期：2017-09-01 00:00:00
abstract：:Operational practice of high-rate anaerobic bioreactors such as upflow anaerobic sludge bed (UASB) reactors is generally based on maximization of the biomass concentration and, in the case of more than one reactor compartment, operation in parallel. In this article, a modeling approach is used to postulate that the tr...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1385/abab:109:1-3:3
更新日期：2003-04-01 00:00:00
abstract：:The increase in waste disposal and energy costs has provided an incentive to convert carbohydrate-rich food waste streams into fuel. For example, dining halls and restaurants discard foods that require tipping fees for removal. An effective use of food waste may be the enzymatic hydrolysis of the waste to simple sugar...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-007-8045-7
更新日期：2008-03-01 00:00:00
abstract：:The authors have recently reported on the design of a protein (MB-1) enriched in methionine, threonine, lysine, and leucine. The protein is intended to be produced by rumen bacteria, in a way that would provide high producing lactating cows with limiting amino acids. In this report, MB-1 stability in the rumen is asse...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/BF02788809
更新日期：1997-04-01 00:00:00
abstract：:A strain of Clostridium thermoaceticum (ATCC 49707) was evaluated for its homoacetate potential. This thermophilic anaerobe best produces acetate from glucose at pH 6.0 and 59 degrees C with a yield of 83% of theoretical. Enzyme hydrolysis of two substrates, a-cellulose and a pulp mill sludge, yielded 68% and 70% dige...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1385/abab:84-86:1-9:963
更新日期：2000-04-01 00:00:00
abstract：:It has been found that lactic acid and hydrogen peroxide (H2O2) displayed co-operatively enhanced killing activity to pathogens. The synergistic effect was investigated with using several microbe species, suggesting that low concentration of lactic acid and H2O2 could kill both Gram-negative and Gram-positive bacteria...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-019-03031-z
更新日期：2019-11-01 00:00:00
abstract：:Pseudomonas are ubiquitous bacteria that can live under a wide range of environmental conditions; some species are successfully used in the fermentation industry. Pseudomonas plecoglossicida JUIM01 strain is an important industrial 2-ketogluconate (2KGA) producer. In Pseudomonas, kguT gene encodes a putative 2KGA tran...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-018-2843-y
更新日期：2019-03-01 00:00:00
abstract：:Sorghum was pretreated by sole ultrasound or supercritical carbon dioxide (scCO2), as well as the method combining both to intensify enzymatic hydrolysis. The effect of the time (1-5 h) and temperature (30-70 °C) on ultrasonic pretreatment was investigated, and the best condition was determined as 5 h and 50 °C with t...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-018-2909-x
更新日期：2019-05-01 00:00:00
abstract：:Proinflammatory cytokines such as tumor necrosis factor (TNF) alpha are well known to inhibit adipocyte differentiation. TNF-alpha triggers ceramide synthesis through binding of TNF-alpha to its p55 receptor. Therefore, ceramide is implicated in many of the multiple signaling pathways initiated by TNF-alpha. In breast...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-008-8505-8
更新日期：2010-01-01 00:00:00
abstract：:Equipment and procedures were developed to study the conversion of lignocellulosic biomass to carboxylic acids using high-solids countercurrent fermentations. Countercurrent fermentations of cattle manure yielded a rapid fermentation (maximum 2.98 g of total acid/[L x d]) with high acid concentrations (maximum of 32.5...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1385/abab:94:2:111
更新日期：2001-05-01 00:00:00
abstract：:The potential carrier role of β-lactoglobulin (β-LG) and its interactions with oxaliplatin were studied using various spectroscopic techniques (fluorescence, UV-visible, and circular dichroism (CD)) in an aqueous medium at two temperatures of 25 and 37 °C in combination with a molecular docking study. Fluorescence mea...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-014-1341-0
更新日期：2015-01-01 00:00:00
abstract：:The recovery of alpha-amylase from the crude enzyme preparation by the reversed micellar liquid-liquid extraction was investigated. The reversed micellar solution was formed by dissolving a cationic surfactant Aliquat 336 in six different alkanes (cyclohexane, n-hexane, isooctane, n-octane, n-decane, and n-dodecane) r...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/BF02787989
更新日期：1997-02-01 00:00:00
abstract：:Foam fractionation is one of the low operating-cost techniques for removing proteins from a dilute solution. The initial bulk solution pH and air superficial velocity play an important role in the foam-fractionation process. Denaturation of proteins (enzymes) can occur, however, during the foam-fractionation process f...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1385/abab:79:1-3:701
更新日期：1999-04-01 00:00:00
abstract：:As a part of our study on microbial heterogeneity in subsurface environments, we have examined the microbial respiration of sediment samples obtained from a coastal site near Oyster, VA. The sediments at the site are unconsolidated, fine to coarse beach sand and gravel. A Columbus Instruments Micro-Oxymax Respirometer...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/BF02941762
更新日期：1996-04-01 00:00:00
abstract：:The bioaugmentation of petroleum-contaminated soil using Enterobacter cloacae was profiled from the evolution of microbial community, soil dehydrogenase activity, to the degradation of petroleum contaminants. The seeding and proliferation of inoculant and the consequential microbial community were monitored by denatur...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-011-9195-1
更新日期：2011-08-01 00:00:00
abstract：:Dissolved oxygen (DO) is an imperative parameter of the activated sludge process (ASP) for wastewater bioremediation. The effect of DO on microbial communities and corresponding metabolic functions in wastewater bioremediation was investigated using next-generation analysis techniques in this study. Illumina-based who...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-020-03340-8
更新日期：2020-10-01 00:00:00
abstract：:Performance of microbial carbon capture cells (MCCs), having a low-cost clayware separator, was evaluated in terms of wastewater treatment and electricity generation using algae Chlorella pyrenoidosa in MCC-1 and Anabaena ambigua in MCC-2 and without algae in a cathodic chamber of MCC-3. Higher power production was ac...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-017-2485-5
更新日期：2017-11-01 00:00:00
abstract：:Gold nanomaterials (Au NMs) have attracted increasing attention in biomedicine due to their facile preparation, multifunctional modifications, unique optical and electrical properties, and good biocompatibility. The physicochemical properties of Au NMs at nanoscale, like size, shape, surface chemistry, and near field ...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章,评审
doi：10.1007/s12010-012-9548-4
更新日期：2012-03-01 00:00:00
abstract：:Tradescantia pallida (Wandering jew)-a succulent perennial herb-was screened to be a potent chromium (Cr) accumulator. Its ability to grow under Cr stress was examined by studying biochemical changes and physiological response of the plant in presence of 5-20 mg L(-1) Cr(VI) concentration in hydroponic environment for...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-014-1035-7
更新日期：2014-08-01 00:00:00
abstract：:Success in generating catalytic antibodies as enzyme mimics lies in the strategic design of the transition-state analog (TSA) for the reaction of interest, and careful development of screening processes for the selection of antibodies that are catalysts. Typically, the choice of TSA structure is straightforward, and t...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1385/abab:83:1-3:195
更新日期：2000-01-01 00:00:00
abstract：:This study investigated methane production in an anaerobic sequencing batch biofilm reactor (AnSBBR) by co-digesting sugarcane vinasse and cheese whey. The assessment was based on the influence of feed strategy, interaction between cycle time and influent concentration, applied volumetric organic load (OLRA), and temp...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-019-03056-4
更新日期：2019-11-01 00:00:00
abstract：:Chitosan/hydroxyapatite composite microparticles were prepared by a solid-in-water-in-oil emulsification cross-linking method. The characteristics and activity in presence of simulated body fluid for 14 and 21 days were investigated. The size distribution, surface morphology, and microstructure of these biomaterials w...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-012-9870-x
更新日期：2012-11-01 00:00:00
abstract：:Neoadjuvant chemotherapy (NACT) is known to be beneficial for patients with locally advanced breast cancer. However, there is still no unified standard on the evaluation of NACT. To identify the potential markers related to NACT sensitivity of breast cancer, in the present study, we examined the protein spectrum of br...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-011-9464-z
更新日期：2012-02-01 00:00:00
abstract：:Lysozymes are known as ubiquitously distributed immune effectors with hydrolytic activity against peptidoglycan, the major bacterial cell wall polymer, to trigger cell lysis. In the present study, the full-length cDNA sequence of a novel sea urchin Strongylocentrotus purpuratus invertebrate-type lysozyme (sp-iLys) was...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-018-2753-z
更新日期：2018-10-01 00:00:00
abstract：:Eighty-nine cultured Pseudomonas species isolated from the sediment and water samples collected from five industrial Red Sea regions that have been affected by petroleum and industry. Genotypic (exoT, exoS, exoU, exoY, lasA, lasB, rhlA, rhlB, Pf1, PAGI-1, -2, and -3) and phenotypic (DNase, elastase, lipase, protease, ...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-017-2553-x
更新日期：2018-01-01 00:00:00
abstract：:Two types of (CNBr-activated) Agarose-staphylococcal endonuclease derivatives have been prepared, one with the enzyme uniformly distributed in the support, and the other with the enzyme preferentially bound in the most external part of the support particles; the latter were obtained using agarose of very small pores a...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/BF02798498
更新日期：1987-02-01 00:00:00
abstract：:A metal oxide semiconductor sensitive to hydrogen was used to monitor H2 production from immobilizedClostridium acetobutylicum cells and from immobilized hydrogenase on a continuous basis. ...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/BF02798635
更新日期：1982-01-01 00:00:00
abstract：:Reed plants (Phragmites communis (Linn.) Trin) are hydrophilic perennial grasses growing in fresh and brackish waters. These plants readily adapt to arid and high salinity conditions; however, their resistance mechanism against abiotic stresses, especially high salinity, is largely unknown. In the present study, we cl...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-015-1514-5
更新日期：2015-04-01 00:00:00
abstract：:The triosephosphate isomerase of Leishmania donovani (LdTIM) was expressed at high level in Escherichia coli. The TIM gene was cloned in expression vector pET-23(a) with C-terminal 6× His tag fused in frame, and expressed as a 27.6-kDa protein in E. coli as inclusion bodies. The recombinant LdTIM from E. coli lysate w...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-011-9206-2
更新日期：2011-08-01 00:00:00
abstract：:Extracellular polymeric substances (EPS) produced by a biofilm-producing bacterium Acinetobacter junii BB1A were characterized. Purified EPS on analysis revealed neutral sugar (73.21 %), uronic acid (10.12 %), amino sugars (0.23 %), α- amino acids (11.13 %), and aromatic amino acids (1.23 %). Infrared spectrometry rev...
journal_title：Applied biochemistry and biotechnology
pub_type： 杂志文章
doi：10.1007/s12010-012-9883-5
更新日期：2012-11-01 00:00:00
