Identification of mobile inhibitors in opposition to Chikungunya virus replication by a cDNA expression cloning mixed with MinION
cDNA expression cloning has been proven to be a strong strategy within the seek for mobile elements that management virus replication. On this examine, cDNA library screening utilizing a pool of cDNA derived from interferon-treated human and porcine cells was mixed with the MinION sequencer to establish mobile genes inhibiting Chikungunya virus (CHIKV) replication.
Problem an infection of CHIKV to Vero cells transduced with the cDNA library produced virus-resistant cells. Then, the MinION sequence of cDNAs extracted from the surviving cells revealed that the open studying frames of TOM7, S100A16, N-terminally truncated type of ECI1 (ECI1ΔN59), and RPL29 have been inserted in lots of the cells.
Importantly, the transient expression of TOM7, S100A16, and ECI1ΔN59 was discovered to inhibit the replication of CHIKV in Huh7 cells, indicating that these mobile elements have been doubtlessly anti-CHIKV molecules.
Thus, our examine demonstrated that cDNA expression cloning mixed with the MinION sequencer allowed a fast and complete detection of mobile inhibitors in opposition to CHIKV.
Description: A polyclonal antibody against XPO6. Recognizes XPO6 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, IHC
Description: A polyclonal antibody against XPO6. Recognizes XPO6 from Human. This antibody is Unconjugated. Tested in the following application: ELISA, IHC; Recommended dilution: IHC:1:20-1:200
Identification of a novel KIR3DL3*064 allele by cDNA cloning and sequencing
Goal: To report on a novel KIR3DL3 allele recognized in a southern Han Chinese language particular person.
Strategies: Peripheral blood pattern was collected from a voluntary blood donor with inconclusive outcome by KIR3DL3 sequence-based typing (SBT). Whole mRNA was extracted and subjected to reverse transcription to acquire KIR3DL3 cDNA, which was then amplified by PCR with a pair of KIR3DL3-specific primers. The product was subjected to cDNA cloning and sequencing.
Outcomes: cDNA cloning and sequencing have recognized a wide-type KIR3DL3*00802 allele and a novel KIR3DL3*064 allele. The latter differed from KIR3DL3*00601 by a missense variant at codon 374[c.1184 C>T (p.Thr374Ile)] in exon 9. The novel KIR3DL3 allele has been formally assigned by the KIR subcommittee of World Well being Group Nomenclature Committee for elements of HLA system.
Conclusion: cDNA cloning and sequencing could also be used to tell apart inconclusive ends in KIR3DL3 SBT with a purpose to establish novel KIR alleles.
Apple Russet Ring and Apple Inexperienced Crinkle Illnesses: Achievement of Koch’s Postulates by Virome Evaluation, Amplification of Full-Size cDNA of Viral Genomes, in vitro Transcription of Infectious Viral RNAs, and Replica of Signs on Fruits of Apple Bushes Inoculated With Viral RNAs
Apple russet ring and apple inexperienced crinkle are graft-transmitted illnesses first reported greater than 60 years in the past, however at current, no affiliation between a selected virus (variant) and the illness has been clearly demonstrated.
On this examine, we carried out the next collection of experiments to establish the causal viruses (variants) of those apple illnesses; (1) complete evaluation by next-generation sequencing of all viruses in every apple tree affected with russet ring or inexperienced crinkle illness,
(2) amplification of full-length genomic cDNA of viruses utilizing primers containing the T3 promoter and the in vitro transcription of infectious viral RNAs, (3) inoculation of viral RNA transcripts to each herbaceous and apple crops, (4) evaluation of sequence variants of viruses current in contaminated crops, (5) back-inoculation of sequence variants of candidate viruses to apple seedlings mixed with the virus-induced flowering know-how utilizing the apple latent spherical virus vector to breed the symptom on the fruit as quickly as doable, and
(6) replica of signs on the fruits of apple timber inoculated with sequence variants and the re-isolation of every virus variant from apples exhibiting fruit signs.
The outcomes confirmed that one of many sequence variants of the apple chlorotic leaf spot virus causes a attribute ring-shaped rust on the fruits of contaminated apple timber and {that a} sequence variant of the apple stem pitting virus in all probability causes inexperienced crinkle signs on an contaminated apple fruit.
Thus, we have been capable of fulfill Koch’s postulates to show the viral etiology of each the apple russet ring and inexperienced crinkle illnesses. We additionally suggest an experimental system that may show whether or not a virus present in diseased tissues is the pathogen liable for the illnesses when the etiology is undetermined.
Identification and characterization of a cDNA encoding a gametocyte-specific protein of the avian coccidial parasite Eimeria necatrix
Gametocyte proteins of Eimeria spp. are essential components of the oocyst wall, and some of these proteins have been analysed to identify targets of transmission-blocking vaccines against avian coccidiosis. In the present study, a cDNA from E. necatrix gametocytes was cloned and sequenced. The cDNA is 1,473 bp in length and encodes a 490-amino-acid protein containing a tyrosine-serine (Tyr/Ser)-rich domain and a proline-methionine (Pro/Met)-rich domain.
A quantitative real-time PCR (qPCR) analysis showed that the cDNA is expressed only during gametogenesis. A fragment containing the Tyr/Ser-rich domain (rEnGAM59) was expressed in Escherichia coli BL21 (DE3) cells. Immunoblotting showed that rEnGAM59 was recognized by the serum of convalescent chickens after infection with E. necatrix, and that an anti-rEnGAM59 antibody recognized a ∼59 kDa protein and two other proteins (∼35 kDa and ∼33 kDa) in gametocyte extracts. An immunofluorescence assay showed that the anti-rEnGAM59 antibody recognized wall-forming bodies in the macrogametocytes and oocyst walls. An in vivo vaccination and challenge trial was conducted to test the potential utility of rEnGAM59 as a vaccine. Immunized chickens performed better than the unimmunized and challenged (positive control) chickens.
The intestinal lesion scores were significantly lower in the immunized groups than in the positive control group (P < 0.05). In contrast, the body weight gains (BWG) were significantly higher in the immunized groups than in the positive control group (P < 0.05). There were no significant differences in the lesion scores and BWG between the groups immunized with rEnGAM59 protein or with live oocysts (P > 0.05).
Chickens immunized with rEnGAM59 protein had a significantly higher antigen-specific serum IgY response (P < 0.05). rEnGAM59 protein can be used as candidate antigen to develop a recombinant coccidiosis vaccine.
Reverse genetics of rotaviruses: Generation of recombinant human rotaviruses from just 11 cDNAs encoding the rotavirus genome
An entirely plasmid-based reverse genetics system for animal rotavirus was established very recently. We improved the reverse genetics system to generate recombinant rotavirus by transfecting only 11 T7 plasmids for its 11 genes under the condition of increasing the ratio (3- or 5-fold) of the cDNA plasmids for NSP2 and NSP5 genes (11-plasmid system).
Utilizing this highly efficient system, we engineered the first infectious recombinant rotaviruses harboring fluorescent (EGFP and mCherry) protein genes. In addition to these recombinant animal viruses, the first infectious recombinant human rotavirus (strain KU (G1P[8])) was also generated with the 11-plasmid system with some modifications.
The availability of recombinant human rotaviruses will provide a genetic platform for a better understanding of the replication, pathogenicity, and other biological characteristics of this medically important virus and enable the rational development of next-generation human rotavirus vaccines.
Lenti ORF clone of Pltp (mGFP-tagged) - Mouse phospholipid transfer protein (Pltp)
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Rat Phospholipid Transfer Protein (PLTP) in serum, plasma, tissue homogenates and other biological fluids.
Rat Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Rat Phospholipid Transfer Protein (PLTP) in serum, plasma, tissue homogenates and other biological fluids.
Rat Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Rat Phospholipid Transfer Protein (PLTP) in serum, plasma, tissue homogenates and other biological fluids.
Rat Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Rat Phospholipid Transfer Protein (PLTP) in serum, plasma, tissue homogenates and other biological fluids.
Rat Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: Enzyme-linked immunosorbent assay based on the Double-antibody Sandwich method for detection of Rat Phospholipid Transfer Protein (PLTP) in samples from Serum, plasma, tissue homogenates and other biological fluids with no significant corss-reactivity with analogues from other species.
Human Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Phospholipid Transfer Protein (PLTP) in serum, plasma, tissue homogenates and other biological fluids.
Human Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Phospholipid Transfer Protein (PLTP) in serum, plasma, tissue homogenates and other biological fluids.
Human Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Phospholipid Transfer Protein (PLTP) in serum, plasma, tissue homogenates and other biological fluids.
Human Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human Phospholipid Transfer Protein (PLTP) in serum, plasma, tissue homogenates and other biological fluids.
Human Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: Enzyme-linked immunosorbent assay based on the Double-antibody Sandwich method for detection of Human Phospholipid Transfer Protein (PLTP) in samples from Serum, plasma, tissue homogenates and other biological fluids with no significant corss-reactivity with analogues from other species.
Mouse Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Phospholipid Transfer Protein (PLTP) in serum, plasma and other biological fluids.
Mouse Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Phospholipid Transfer Protein (PLTP) in serum, plasma and other biological fluids.
Mouse Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Phospholipid Transfer Protein (PLTP) in serum, plasma and other biological fluids.
Mouse Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Phospholipid Transfer Protein (PLTP) in serum, plasma and other biological fluids.
Mouse Phospholipid Transfer Protein (PLTP) ELISA Kit
Description: Enzyme-linked immunosorbent assay based on the Double-antibody Sandwich method for detection of Mouse Phospholipid Transfer Protein (PLTP) in samples from Serum, plasma and other biological fluids with no significant corss-reactivity with analogues from other species.
Polyclonal Antibody to Phospholipid Transfer Protein (PLTP)
Description: A Monoclonal antibody against Human PLTP (monoclonal) (M01). The antibodies are raised in mouse and are from clone 2F3-G4. This antibody is applicable in WB and IHC
Phospholipid Transfer Protein (PLTP) Polyclonal Antibody (Mouse), FITC
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 - Mouse exportin 6 (Xpo6))
 - Mouse exportin 6 (Xpo6))
 - Rat exportin 6 (Xpo6), (10 ug))
 - Rat exportin 6 (Xpo6), (10 ug))
, mGFP tagged)
 for miRNA target validation)
, Myc-DDK-tagged)
-Human exportin 6 (XPO6))
 - Human exportin 6 (XPO6))
 - Mouse exportin 6 (Xpo6), (10ug))
 - Mouse phospholipid transfer protein (Pltp))
)
 - Mouse phospholipid transfer protein (Pltp))
)
 ELISA Kit)
 ELISA Kit)
 ELISA Kit)
)
 Polyclonal Antibody (Rat))
 Polyclonal Antibody (Mouse))
 Polyclonal Antibody (Rat), PE)
 Polyclonal Antibody (Rat), APC)
 Polyclonal Antibody (Rat), Cy3)
 Polyclonal Antibody (Rat), HRP)
 Polyclonal Antibody (Mouse), APC)
 Polyclonal Antibody (Mouse), Cy3)
 Polyclonal Antibody (Mouse), HRP)
 Polyclonal Antibody (Mouse), PE)
 Polyclonal Antibody (Rat), FITC)
 (M01), Clone: 2F3-G4)
 Polyclonal Antibody (Mouse), FITC)
 Polyclonal Antibody (Rat), Biotinylated)
 Polyclonal Antibody (Mouse), Biotinylated)
 Polyclonal Antibody (Rat), APC-Cy7)
 Polyclonal Antibody (Mouse), APC-Cy7)
