scholarly journals Study of the 49 kDa excretory-secretory protein gene of Trichinella nativa and Trichinella spiralis

2007 ◽  
Vol 44 (3) ◽  
pp. 120-125 ◽  
Author(s):  
B. Zheng ◽  
L. Xiao ◽  
X. Wang ◽  
D. Li ◽  
Y. Lu ◽  
...  
Keyword(s):  
Prokaryotic Expression ◽  
Protein Gene ◽  
Trichinella Spiralis ◽  
Secretory Protein ◽  
Rt Pcr ◽  
Prokaryotic Expression Vector ◽  
Close Relationship ◽  
Bamhi Site ◽  
Relationship Of ◽  
Trichinella Nativa

AbstractTo study the function of the 49 kDa excretory-secretory (ES) protein gene (P49) of Trichinella, the genes was amplified by RT-PCR from RNA of Trichinella spiralis and Trichinella nativa and several Chinese Trichinella isolates of domestic animals, and sequenced after being cloned. The amplified products of these parasites produced bands of about 950 bp. The 97.2 % to 100 % nucleotides identity and 94.3 % to 100 % identity of deduced amino acids among P49 gene of these Trichinella strains showed the close relationship of these parasites. The P49 gene of T. nativa was cloned into the BamHI site of the prokaryotic expression vector pET-30a, and the recombinant vector was expressed. The expressed product was 40.8 kDa in size. In Western blot analysis, the expressed product was reactive to sera of mice infected with T. nativa, T. spiralis and their Chinese geographical strains.

Prokaryotic Expression and Purification of Tomato Cnr Protein

2013 ◽  
Vol 690-693 ◽  
pp. 1247-1251
Author(s):  
Yuan Hong Xie ◽  
Hui Liu ◽  
Hong Xing Zhang
Keyword(s):  
Fruit Ripening ◽  
Prokaryotic Expression ◽  
Autosomal Dominant ◽  
Tomato Fruit ◽  
Rt Pcr ◽  
Expression And Purification ◽  
Tomato Fruit Ripening ◽  
Prokaryotic Expression Vector ◽  
Dominant Mutant ◽  
Natural Mutation

Tomato fruit ripening mutant colorless non-ripening (Cnr) is an autosomal dominant mutant, obtained by natural mutation. The fruit of Cnr mutant was obviously immature, and the softening and lycopene accumulation of the Cnr fruit was significantly inhibited during the fruit ripening. In this work, we cloned the Cnr gene from the red ripe tomato fruit by RT-PCR, and constructed a prokaryotic expression vector. And the recombinant Cnr protein was induced by 1 mM IPTG in Escherichia coli BL21 (DE3), and purified by MBP affinity chromatography.

scholarly journals MOLECULAR CLONING, PROKARYOTIC EXPRESSION AND PREPARATION OF ANTISERUM AGAINST LILY SYMPTOMLESS VIRUS TGB1 PROTEIN

2017 ◽  
Vol 7 (1) ◽  
pp. 986-990
Author(s):  
Pinsan Xu ◽  
Xinlei Wang ◽  
Zhengyao Zhang
Keyword(s):  
Prokaryotic Expression ◽  
Triple Gene Block ◽  
Rapid Test ◽  
Coli Strain ◽  
Functional Study ◽  
Rt Pcr ◽  
Lily Symptomless Virus ◽  
Gene Block ◽  
Prokaryotic Expression Vector ◽  
Preparation Of Antiserum

 The triple gene block gene TGB1 was amplified by RT-PCR from lily leaves infected with Lily symptomless virus and cloned into prokaryotic expression vector pET-28a(+). The recombinant vector was transformed into Escherichia coli strain BL21 (DE3). On induction with isopropyl β-D-1-thiogalactopyranoside,TGB1 protein was highly expressed and the molecular weight was 29 kDa (including a His-tag-containing fusion). After protein purification by Ni2+-NTA affinity chromatography, a polyclonal antibody against TGB1 was raised in mouse. Western blot analysis showed that the antiserum reacted specially with the TGB1 protein of LSV. ELISA and RT-PCR confirmed that the antiserum reacted specially with lily leaves infected with LSV, and can be used for a rapid test for LSV. The antibody produced in this work may be used for future immunohistochemistry and functional study of TGB1.

scholarly journals Chayote mosaic virus, a New Tymovirus Infecting Cucurbitaceae

2000 ◽  
Vol 90 (10) ◽  
pp. 1098-1104 ◽  
Author(s):  
Juan J. Bernal ◽  
Ignacio Jiménez ◽  
Manuel Moreno ◽  
Melanie Hord ◽  
Carmen Rivera ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Protein Gene ◽  
Phylogenetic Analyses ◽  
Disease Symptoms ◽  
Close Relationship ◽  
Andean Potato ◽  
The Caribbean ◽  
Asymptomatic Infections ◽  
Relationship Of ◽  
Host Families

Chayote mosaic virus (ChMV) is a putative tymovirus isolated from chayote crops in Costa Rica. ChMV was characterized at the host range, serological, and molecular levels. ChMV was transmitted mechanically and induced disease symptoms mainly in Cucurbitaceae hosts. Asymptomatic infections were detected in other host families. Serologically, ChMV is related to the Andean potato latent virus (APLV) and the Eggplant mosaic virus (EMV), both members of the genus Tymovirus infecting solanaceous hosts in the Caribbean Basin and South America. The sequence of the genomic RNA of ChMV was determined and its genetic organization was typical of tymoviruses. Comparisons with other tymoviral sequences showed that ChMV was a new member of the genus Tymovirus. The phylogenetic analyses of the coat protein gene were consistent with serological comparisons and positioned ChMV within a cluster of tymoviruses infecting mainly cucurbit or solanaceous hosts, including APLV and EMV. Phylogenetic analyses of the replicase protein gene confirmed the close relationship of ChMV and EMV. Our results suggest that ChMV is related to two tymoviruses (APLV and EMV) of proximal geographical provenance but with different natural host ranges. ChMV is the first cucurbit-infecting tymovirus to be fully characterized at the genomic level.

PROKARYOTIC EXPRESSION OF RECOMBINANT MOUSE OP-1 AND ITS OSTEOGENESIS IN VITRO

2002 ◽  
Vol 06 (01) ◽  
pp. 9-15
Author(s):  
Zhenhua Lu ◽  
T. Sam Lindholm
Keyword(s):  
Recombinant Protein ◽  
Prokaryotic Expression ◽  
Development Research ◽  
Activity Assay ◽  
Rt Pcr ◽  
Iptg Induction ◽  
Reading Frame ◽  
Prokaryotic Expression Vector ◽  
Osteogenesis In Vitro

OP-1 is a main member of the BMP family. It plays diverse and significant roles in growth and differentiation. RT-PCR amplified the open reading frame of mouse OP-1 from cDNAs synthesis from CD-1 mouse embryo. By coding the gene of mOP-1 was inserted into pTrcHis 2B, the prokaryotic expression vector through IPTG induction, and the recombinant protein was isolated and purified with Ni-NTA resin. In an ALP activity assay, rmOP-1 has shown osteogenesis activity in vitro. It would offer a fast and inexpensive means of the productions of recombinant protein and an easy tool for growth and development research on rmOP-1.

NOTES ON THE GENUS PROTARCHOIDES (HYMEN., ICHNEUMONIDAE)

1938 ◽  
Vol 70 (11) ◽  
pp. 230-232 ◽  
Author(s):  
C. Stuart Walley
Keyword(s):  
North American ◽  
North American Species ◽  
Close Relationship ◽  
Relationship Of

The following notes were assembled in arranging the Protarchoides material in the National Collection. In establishing the identity of Protarchoides mellipes (Prov.) it has been found necessary to synonymize one species. A species allied to mellipes is described as new and a table is provided for the separation of the four known North American species. The recording of Trichiosoma as host for a member of this genus is further evidence of the close relationship of the genus with Protarchus Foer.

Actinomycetes as the basis of Probiotics for Plants

2021 ◽  
Vol 37 (2) ◽  
pp. 3-19
Author(s):  
A.A. Gagarina
Keyword(s):  
Plant Growth ◽  
Wide Distribution ◽  
Present Review ◽  
Rhizosphere Microorganisms ◽  
Current State ◽  
Close Relationship ◽  
Minimum Requirements ◽  
Stimulate Plant Growth ◽  
Relationship Of ◽  
Streptomyces Genus

The present review describes the concept of probiotics for plants and analyzes the prospects for using actinomycetes as producers of these drugs. The minimum requirements for plant probiotic microorganisms are proposed, similar to those for human probiotic microorganisms. These are utility, efficiency and safety for plants, as well as mandatory isolation from plant samples. It is noted that these requirements are usually met by endophytic and rhizosphere microorganisms that stimulate plant growth and provide them with protection from phytopathogens. Evidence is given for the possibility of attributing actinomycetes to probiotic plant bacteria, due to the close relationship of these microorganisms with plants, their wide distribution in populations of endophytic and rhizosphere microorganisms, and the presence of phytoregulatory activity. The review provides examples of genera and species of actinomycetes that are promising producers of probiotics for agronomically important crops. The most studied and commercialized of them are representatives of the Streptomyces genus. The current state, prospects and problems in commercialization of probiotics based on actinomycetes are discussed. probiotic microorganisms of plants, associative actinomycetes, endophytes, rhizosphere, biological preparations

Identification ofTrichinella spiralisearly antigens at the pre-adult and adult stages

Parasitology ◽  
2010 ◽  
Vol 138 (4) ◽  
pp. 463-471 ◽  
Author(s):  
ALEKSANDAR ZOCEVIC ◽  
PAULINE MACE ◽  
ISABELLE VALLEE ◽  
RADU BLAGA ◽  
MINGYUAN LIU ◽  
...  
Keyword(s):  
Cdna Library ◽  
Serine Proteases ◽  
Trichinella Spiralis ◽  
Serine Proteinase ◽  
Secretory Protein ◽  
Cdna Libraries ◽  
Sequence Identity ◽  
Muscle Larvae ◽  
Serine Protease Family ◽  
Host Parasite

SUMMARYThree expression cDNA libraries fromTrichinella spiralisworms 14 h, 20 h and 48 h post-infection (p.i.) were screened with serum from pigs experimentally infected with 20 000T. spiralismuscle larvae. Twenty-nine positive clones were isolated from the 14 h p.i. cDNA library, corresponding to 8 different genes. A putative excretory-secretory protein similar to that ofT. pseudospiraliswas identified. Three clones corresponded to aT. spiralisserine proteinase inhibitor known to be involved in diverse functions such as blood coagulation and modulation of inflammation. Screening of the 20 h p.i. cDNA library selected 167 positive clones representing 12 different sequences. The clone with the highest redundancy encoded a small polypeptide having no sequence identity with any known proteins fromTrichinellaor other organisms. Fourteen clones displayed sequence identity with the heat shock protein (HSP) 70. HSPs are produced as an adaptive response of the parasite to the hostile environment encountered in the host intestine but their mechanism of action is not yet well defined. From the 48 h p.i.T. spiraliscDNA library, 91 positive clones were identified representing 7 distinct sequences. Most of the positive clones showed high similarity with a member of a putativeT. spiralisserine protease family. This result is consistent with a possible major role for serine proteases during invasive stages ofTrichinellainfection and host-parasite interactions.

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