Paléontologie Moleculaire, PCR, mRNA, cDNA, miRNA, Total RNA
Identification and characterization of a cDNA encoding a gametocyte-specific
JuanJune 25, 20210 Comments
Identification of salinity responsive genes in lavender by the use of cDNA-AFLP
Presently, a world demand exists forlavender as an enormous medicinal plant and provide of essential oils. Freshwater and arable lands are two predominant parts that inhibit intensive farming of medicinal crops in Iran. Saline water from seas and salty soil is also new sources for agricultural use, significantly for medicinal crops.
We sought to extend our information of the Lavandulaangustifolia genome and molecular basis of its salinity tolerance by way of using cDNA amplified fragment measurement polymorphism (cDNA-AFLP) to research the changes in plant transcriptomes in response to NaCl.
All acknowledged transcript derived fragments (TDF) have been assigned as novel angustifolia genes related to signal transduction, regulation of gene expression, totally different splicing, autophagy, and secondary metabolite biosynthesis.
qRT-PCR analysis of the TDFs in response to fully totally different concentrations of NaCl revealed assorted ranges of mRNA of the acknowledged genes on this plant.
Our findings provided predominant insights into the molecular response of angustifolia to salinity.
Description: A DNA sequence encoding the mature variant of ov-VEGF-E isolate D1701 (Dehio et al., 1999; GenBank accession No. AF106020) was expressed in E. coli as a 132 amino acid residue fusion protein with an N-terminal His-tag sequence and a thrombin cleavage site. Recombinant VEGF-E homodimer was dimerized in vitro and has a predicted mass of approximately 35 kDa. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV) (Lyttle et al., 1994). Different isolates of Orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appears to be derived from captured host genes. All eight Cysteine residues of the central Cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins (Dehio et al., 1999; Wise et al., 1999). Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E cannot bind to VEGF receptor-1 (Flt-1). Furthermore VEGF-E can also not bind to VEGF receptor-3 (FLT-4). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor –2/KDR.
Description: A DNA sequence encoding the mature variant of ovVEGF-E isolate D1701 (Dehio et al., 1999; GenBank accession No. AF106020) was expressed in E. coli as a 132 amino acid residue fusion protein with an N-terminal His-tag sequence and a thrombin cleavage site. Recombinant VEGF-E homodimer was dimerized in vitro and has a predicted mass of approximately 35 kDa. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV) (Lyttle et al., 1994). Different isolates of Orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appears to be derived from captured host genes. All eight cysteine residues of the central cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins (Dehio et al., 1999; Wise et al., 1999). Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E can not bind to VEGF receptor-1 (Flt-1). Furthermore VEGF-E can also not bind to VEGF receptor-3 (FLT-4). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor –2/KDR.
Description: A DNA sequence encoding the mature variant of ovVEGF-E isolate D1701 (Dehio et al., 1999; GenBank accession No. AF106020) was expressed in E. coli as a 132 amino acid residue fusion protein with an N-terminal His-tag sequence and a thrombin cleavage site. Recombinant VEGF-E homodimer was dimerized in vitro and has a predicted mass of approximately 35 kDa. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV) (Lyttle et al., 1994). Different isolates of Orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appears to be derived from captured host genes. All eight cysteine residues of the central cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins (Dehio et al., 1999; Wise et al., 1999). Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E can not bind to VEGF receptor-1 (Flt-1). Furthermore VEGF-E can also not bind to VEGF receptor-3 (FLT-4). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor –2/KDR.
Description: A DNA sequence encoding the mature variant of ov-VEGF-E isolate D1701 was expressed in E. coli as a 132 amino acid residue fusion protein with an N-terminal His-tag sequence and a thrombin cleavage site. Recombinant VEGF-E homodimer was dimerized in vitro and has a predicted mass of approximately 35 kDa. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV). Different isolates of Orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appears to be derived from captured host genes. All eight Cysteine residues of the central Cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins. Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E cannot bind to VEGF receptor-1 (Flt-1). Furthermore VEGF-E can also not bind to VEGF receptor-3 (FLT-4). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor –2/KDR.
Recombinant Virus VEGF E (Orf Virus) Protein, His, E.coli-1mg
Description: A DNA sequence encoding the first 116 amino acid residue of Orf virus VEGF-E isolate D1701 (Dehio et al., 1999 EMBO J. 18:363-374; GenBank accession No. AF106020) was fused with a DNA sequence encoding to the C-terminal heparin binding domain of human VEGF165. The chimeric protein was expressed in insect cells using a baculovirus expression system. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV) (Lyttle et al., 1994 J. Virol 68:84-92). Different isolates of orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appear to be derived from captured host genes. All eight cysteine residues of the central cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins (Dehio et al., 1999 EMBO J. 18:363-374; Wise et al., 1999 Proc. Natl. Acad. Sci USA 96:3071-3076). Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E and hb-VEGF-E can not bind to VEGF receptor-1 (Flt-1). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor–2/ KDR. Compared to human VEGF165 this virus form has no heparin-binding domain and seems to be a freely secreted protein comparable to VEGF121. In order to compare this form with human VEGF165, an additional heparin-binding domain was engineered at the C-terminus to allow interaction with proteo-aminoglycans and heparan sulfate. These form is also able to interact with neuropillin–1.
Orf virus VEGF-E, Heparin-binding Recombinant Protein
Description: A DNA sequence encoding the first 116 amino acid residue of Orf virus VEGF-E isolate D1701 (Dehio et al., 1999 EMBO J. 18:363-374; GenBank accession No. AF106020) was fused with a DNA sequence encoding to the C-terminal heparin binding domain of human VEGF165. The chimeric protein was expressed in insect cells using a baculovirus expression system. Based on sequence similarity to VEGF-A, a gene encoding a VEGF homologue has recently been discovered in the genome of Orf virus (OV) (Lyttle et al., 1994 J. Virol 68:84-92). Different isolates of orf virus show significant amino acid sequence similarity to VEGF-A and described as a viral virulence factor that appear to be derived from captured host genes. All eight cysteine residues of the central cysteine knot motif characteristic of members of the VEGF family are conserved among other residues in the VEGF-E proteins (Dehio et al., 1999 EMBO J. 18:363-374; Wise et al., 1999 Proc. Natl. Acad. Sci USA 96:3071-3076). Alignment of all mammalian VEGF sequences indicated that VEGF-E is distinct from the previously described VEGFs but most closely related to VEGF-A. Like VEGF-A, VEGF-E was found to bind with high affinity to VEGF receptor-2 (KDR) resulting in receptor autophosphorylation, whilst in contrast to VEGF-A, VEGF-E and hb-VEGF-E can not bind to VEGF receptor-1 (Flt-1). Therefore VEGF-E is a potent angiogenic factor selectively binding to VEGF receptor–2/ KDR. Compared to human VEGF165 this virus form has no heparin-binding domain and seems to be a freely secreted protein comparable to VEGF121. In order to compare this form with human VEGF165, an additional heparin-binding domain was engineered at the C-terminus to allow interaction with proteo-aminoglycans and heparan sulfate. These form is also able to interact with neuropillin–1.
Description: A DNA sequence encoding the mature variant of ovVEGF-E isolate D1701 (Dehio et al., 1999; GenBank accession No. AF106020) was expressed in E. coli as a 132 amino acid residue fusion protein with an N-terminal His-tag sequence and a thrombin cleavage site. Recombinant VEGF-E homodimer was dimerized in vitro and has a predicted mass of approximately 35 kDa.
Identification and characterization of a cDNA encoding a gametocyte-specific protein of the avian coccidial parasite Eimerianecatrix
Gametocyte proteins of Eimeria spp. are essential parts of the oocyst wall, and some of those proteins have been analysed to determine targets of transmission-blocking vaccines in opposition to avian coccidiosis. Throughout the present look at, a cDNA from E. necatrix gametocytes was cloned and sequenced. The cDNA is 1,473 bp in measurement and encodes a 490-amino-acid protein containing a tyrosine-serine (Tyr/Ser)-rich space and a proline-methionine (Skilled/Met)-rich space.
A quantitative real-time PCR (qPCR) analysis confirmed that the cDNA is expressed solely all through gametogenesis. A fraction containing the Tyr/Ser-rich space (rEnGAM59) was expressed in Escherichia coli BL21 (DE3) cells. Immunoblotting confirmed that rEnGAM59 was acknowledged by the serum of convalescent chickens after an an infection with E. necatrix, and that an anti-rEnGAM59 antibody acknowledged a ∼59 kDa protein andtwo totally different proteins (∼35 kDa and ∼33 kDa) in gametocyte extracts.
An immunofluorescence assay confirmed that the anti-rEnGAM59 antibody acknowledged wall-forming our our bodies throughout the macrogametocytes and oocyst partitions. An in vivo vaccination and drawback trial was carried out to verify the potential utility of rEnGAM59 as a vaccine.
Immunized chickens carried out increased than the unimmunized and challenged (constructive administration) chickens. The intestinal lesion scores have been significantly lower throughout the immunized groups than throughout the constructive administration group (P < 0.05). In distinction, the physique weight optimistic elements (BWG) have been significantly bigger throughout the immunized groups than throughout the constructive administration group (P < 0.05). There have been no very important variations throughout the lesion scores and BWG between the groups immunized with rEnGAM59 protein or with keep oocysts (P > 0.05). Chickens immunized with rEnGAM59 protein had a significantly bigger antigen-specific serum IgY response (P < 0.05). rEnGAM59 protein may be utilized as candidate antigen to develop a recombinant coccidiosis vaccine.
Reverse genetics of rotaviruses: Era of recombinant human rotaviruses from simply 11 cDNAs encoding the rotavirus genome
A very plasmid-based reverse genetics system for animal rotavirus was established very these days. We improved the reverse genetics system to generate recombinant rotavirus by transfecting solely 11 T7 plasmids for its 11 genes beneath the scenario of accelerating the ratio (3- or 5-fold) of the cDNA plasmids for NSP2 and NSP5 genes (11-plasmid system).
Utilizing this extraordinarily surroundings pleasant system, we engineered the first infectious recombinant rotaviruses harboring fluorescent (EGFP and mCherry) protein genes. Together with these recombinant animal viruses, the first infectious recombinant human rotavirus (stress KU (G1P[8])) was moreover generated with the 11-plasmid system with some modifications.
The availability of recombinant human rotaviruses will current a genetic platform for a higher understanding of the replication, pathogenicity, and totally different natural traits of this medically very important virus and permit the rational enchancment of next-generation human rotavirus vaccines.
Protein expression-based classification of gastric most cancers by immunohistochemistry of tissue microarray
Lately, the Most cancers Genome Atlas and Asian Most cancers Analysis Group suggest two new classifications system of gastric most cancers through the use of multi-platforms of molecular analyses. Nevertheless, these extremely difficult and price applied sciences haven’t but been translated into full medical utility. As well as, the clinicians are anticipated to realize extra steerage of therapy for various molecular subtypes.
On this research, we developed a panel of gastric most cancers sufferers in inhabitants from Southern China utilizing commercially accessible TMA and immunohistochemical know-how. A cohort of 259 GC sufferers was labeled into four subtypes on the premise of expression of mismatch restore proteins (PMS2, MLH1, MSH2, and MSH6), E-cadherin and p21 protein. We noticed that the subtypes offered distinct prognosis.
dMMR-like subtype was related to the perfect prognosis, and E-cadherin-a subtype was related to the worst prognosis. Sufferers with p21-Excessive and p21-Ligh subtypes had intermediate general survival. In multivariate evaluation, the dMMR-like subtype remained an impartial prediction energy for general survival within the mannequin. We described a molecular classification of gastric cancers utilizing clinically relevant assay.
The organic relevance of the 4 subtypes was illustrated by vital variations in prognosis. Our molecular classification offered an efficient and cheap screening software for bettering prognostic fashions. Nonetheless, our research needs to be thought of preliminary and carries a restricted predictive worth as a single-center retrospective research.
Description: This gene encodes complement factor B, a component of the alternative pathway of complement activation. Factor B circulates in the blood as a single chain polypeptide. Upon activation of the alternative pathway, it is cleaved by complement factor D yielding the noncatalytic chain Ba and the catalytic subunit Bb. The active subunit Bb is a serine protease which associates with C3b to form the alternative pathway C3 convertase. Bb is involved in the proliferation of preactivated B lymphocytes, while Ba inhibits their proliferation. This gene localizes to the major histocompatibility complex (MHC) class III region on chromosome 6. This cluster includes several genes involved in regulation of the immune reaction. Polymorphisms in this gene are associated with a reduced risk of age-related macular degeneration. The polyadenylation site of this gene is 421 bp from the 5' end of the gene for complement component 2.
Description: This gene encodes complement factor B, a component of the alternative pathway of complement activation. Factor B circulates in the blood as a single chain polypeptide. Upon activation of the alternative pathway, it is cleaved by complement factor D yielding the noncatalytic chain Ba and the catalytic subunit Bb. The active subunit Bb is a serine protease which associates with C3b to form the alternative pathway C3 convertase. Bb is involved in the proliferation of preactivated B lymphocytes, while Ba inhibits their proliferation. This gene localizes to the major histocompatibility complex (MHC) class III region on chromosome 6. This cluster includes several genes involved in regulation of the immune reaction. Polymorphisms in this gene are associated with a reduced risk of age-related macular degeneration. The polyadenylation site of this gene is 421 bp from the 5' end of the gene for complement component 2.
Description: This gene encodes complement factor B, a component of the alternative pathway of complement activation. Factor B circulates in the blood as a single chain polypeptide. Upon activation of the alternative pathway, it is cleaved by complement factor D yielding the noncatalytic chain Ba and the catalytic subunit Bb. The active subunit Bb is a serine protease which associates with C3b to form the alternative pathway C3 convertase. Bb is involved in the proliferation of preactivated B lymphocytes, while Ba inhibits their proliferation. This gene localizes to the major histocompatibility complex (MHC) class III region on chromosome 6. This cluster includes several genes involved in regulation of the immune reaction. Polymorphisms in this gene are associated with a reduced risk of age-related macular degeneration. The polyadenylation site of this gene is 421 bp from the 5' end of the gene for complement component 2.
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human CFB (Center). This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody raised in Goat that recognizes and binds to Human CFB / Complement Factor B . This antibody is tested and proven to work in the following applications:
