ABSTRACT
Idiopathic pulmonary fibrosis （IPF） is a common， lethal interstitial lung disease characterized by airway remodeling， inflammation， alveolar destruction， and fibrosis. The mammalian target of rapamycin complex 1/4E binding protein 1 （mTORC1/4E-BP1） axis is closely related to the expression of collagen by fibroblasts， and its role in pulmonary fibrosis remains to be further elucidated. Traditional Chinese medicine （TCM） has shown promising efficacy in improving the lung function， exercise capacity， and quality of life in patients with IPF. The theory of "same treatment for different diseases" provides a TCM theoretical basis for the treatment of pulmonary fibrosis with Bupleuri Radix， while the research in western medicine has preliminarily shown that both the formulation and single herb as well as the active ingredients of Bupleuri Radix have good therapeutic effects on pulmonary fibrosis. Therefore， this review will elaborate on the role of the mTORC1/4E-BP1 axis in the pathomechanism of IPF， as well as the research results of the active components of Bupleuri Radix on the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin protein（PI3K/AKT/mTOR） pathway， so as to provide a reference for the treatment and drug development of IPF.
ABSTRACT
24-Alkyl sterols are the major players in the control of membrane component and plant growth. In this paper, we cloned an important rate-limiting enzyme:sterol-C-24-methyl transferase (SMT) in the sterol biosynthetic pathway according to the transcriptome data of Tripterygium wilfordii. suspension cells, whose full-length cDNA was 1 631 bp with an open reading frame of 1 080 bp, encoding a protein of 359 amino acids. It was estimated that theoretical isoelectric point (pI) was 6.43 and the molecular mass was 40.0 kDa. Bioinformatics analysis attributed the SMT gene to SMT2 family. The expression vector was constructed as the pMAL-c2x-TwSMT2 plasmid and the recombinant protein was expressed in E. coil BL21 (DE3) competent cells. After methyl jasmonate treatment, the relative expression level of TwSMT2 has improved significantly in 24 h. SDS-PAGE electrophoresis and Western Blot showed that protein of TwSMT2 in BL21 (DE3) strain was expressed after induction by IPTG. In this study, TwSMT2 was cloned for the first time and the recombinant protein was expressed, which lay the foundation for elucidation of the sterol biosynthetic pathway of Tripterygium wilfordii in the future.
ABSTRACT
In this paper, we cloned the full-length cDNA of TwSQS from Tripterygium wilfordii suspension cells (GenBank:KR401220) and performed the bioinformation and mRNA expression analysis. The expression after methyl jasmonate (MJ) treatment of the gene was detected by RT-PCR. The full-length cDNA of TwSQS was 1800 bp containing a 1242 bp open reading frame (ORF) encoding a polypeptide of 413 amino acids. The theoretical isoelectric point (pI) was 7.94 and the calculate molecular weight was about 47.20 kD. The relative expression level of TwSQS was deduced by MJ and reached the highest at 4 h after the treatment. The gene information we got in this study enriched the biosynthesis pathway of triterpenoids in Tripterygium wilfordii and laid foundation for further studies.
ABSTRACT
A full-length cDNA of GGPPS gene from Tripterygium wilfordii suspension cells was obtained by use of RACE strategy (GeneBank: KM978333), and then analyzed by bioinformatics approaches. TwGGPPS cDNA has 1857 nucleotides and an open reading frame (ORF) encoding a protein of 514 amino acid residues. The deduced protein has isoelectric point (pI) of 7.85, a calculated molecular weight about 57.13 kD, 5 conserved domains and 2 functional domains. PSORT Prediction showed it was located at plasma membrane. Phylogenetic analysis demonstrated that TwGGPPS1 was similar to GGPPS from other species of plants. For the first time the cloning of geranylgeranyl diphosphate synthase gene from T. wilfordii was reported, it lays the foundation for further research of diterpenoids biosynthetic pathway.
Subject(s)
Amino Acid Sequence , Cloning, Molecular , Farnesyltranstransferase , Chemistry , Genetics , Metabolism , Molecular Sequence Data , Phylogeny , Plant Proteins , Chemistry , Genetics , Metabolism , Sequence Alignment , Sequence Homology, Amino Acid , Tripterygium , Chemistry , GeneticsABSTRACT
In this study, based on the transcriptome data, we cloned the full-length cDNAs of TwAACT gene from Tripterygium wilfordii suspension cells, and then analyzed the bioinformation of the sequence, detected the genetic differential expression after being induced by methyl jasmonate (MeJA) by RT-PCR. The full-length cDNA of the TwAACT was 1 704 bp containing a 1 218 bp open reading frame (ORF) encoding a polypeptide of 405 amino acids (GeneBank accession No. KP297934). The deduced isoelectric point (pI) was 6.10, a calculated molecular weight was about 41.20 kDa, and online prediction showed that TwAACT had two catalytic active sites. After the induction of MeJA, the relative expression level of TwAACT increased rapidly. The expression level of TwAACT was highest at 24 h. TwAACT was cloned firstly, that laid the foundation for identifying thegene and illustrating thebiosynthesis mechanism and its synthetic biology.
Subject(s)
Acetyl-CoA C-Acetyltransferase , Chemistry , Genetics , Metabolism , Amino Acid Sequence , Cloning, Molecular , Gene Expression Regulation, Plant , Models, Molecular , Molecular Sequence Data , Phylogeny , Plant Proteins , Chemistry , Genetics , Metabolism , Sequence Alignment , Tripterygium , Chemistry , Classification , GeneticsABSTRACT
Based on the transcriptome database of Salvia miltiorrhiza, specific primers were designed to clone a full-length cDNA of ent-kaurene oxidase synthase (SmKOL) using the RACE strategy. ORF Finder was used to find the open reading frame of SmKOL cDNA, and ClustalW has been performed to analysis the multiple amino acid sequence alignment. Phylogenetic tree has been constructed using MEGA 5.1. The transcription level of SmKOL from the hairy roots induced by elicitor methyl jasmonate (MeJA) was qualifiedby real-time quantitative PCR. The full length of SmKOL cDNA was of 1 884 bp nucleotides encoding 519 amino acids. The molecular weight of the SmKOL protein was about 58.88 kDa with isoelectric point (pI) of 7.62. Results of real-time quantitative PCR analyses indicated that the level of SmKOL mRNA expression in hairy roots was increased by elicitor oMeJA, and reached maximum in 36 h. The full-length cDNA of SmKOL was cloned from S. miltiorrhiza hairy root, which provides a target gene for further studies of its function, gibberellin biosynthesis and regulation of secondary metabolites.
Subject(s)
Amino Acid Sequence , Cloning, Molecular , Computational Biology , Methods , Cytochrome P-450 Enzyme System , Chemistry , Genetics , Models, Molecular , Molecular Sequence Data , Phylogeny , Protein Structure, Tertiary , Salvia miltiorrhizaABSTRACT
The aim of this study was to establishment of known HPA genotype platelet donor data base and carry out the platelet antigen gene typing and matching in the platelet transfusion so as to prevent the production of the antibody and to avoid the platelet transfusion refractoriness. HPA of the donors and the patients were genotyped by PCR-SSP. The platelet transfusion with the same ABO type and HPA genotypes as the patient was selected and infused if the solid-phase agglutination matching was consistent. The results showed that the distribution of HPA types in Chinese Mudanjiang area had its own characteristics. HPA-15 had the greatest heterozygosity, and then HPA-3. The effective rate was 94.4% if both the HPA types and the ABO types were coincident while the effective rate was 77.8% when only ABO types were coincident. It is concluded that the distribution of HPA types in Chinese Mudanjiang area presents genetic polymorphism. The platelet transfusion with genetic typing and matching of both HPA and ABO types is an effective way to avoid production of the antibody and platelet transfusion refractoriness.
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Antigens, Human Platelet , Genetics , Allergy and Immunology , Blood Grouping and Crossmatching , Genotype , Platelet Transfusion , MethodsABSTRACT
There exists many kinds and a huge number of terpenoid in medicinal plants, which show a wide range of pharmacological activities. 3-Hydroxy-3-metllylglutaryl coenzyme A reductase(HMGR) is a key rate-limiting enzyme in terpenoid biosynthetic pathway . HMGR plays an important role in the regulation of secondary metabolism of the terpenoid. The paper summarized the biological function and the catalytic mechanism of HMGR, the cloning and the structure of the gene as well as its research progress in some medicinal plants.
Subject(s)
Hydroxymethylglutaryl CoA Reductases , Metabolism , Plants, Medicinal , Terpenes , MetabolismABSTRACT
A series of experiments were conducted to study the mutation of Kloechera apiculata by many kinds of treatments such as UV and UV+LiCl.The optimal dosage disposal was determined:15 W 30 cm under Ultraviolet irradiation for 20 s,UV+LiCl under Ultraviolet irradiation for 20 s and added LiCl 0.3% (w/v).One strain(UV20-13)which had obvious physiological characteristic was obtained.the incidence of blue and green mold of citrus was reduced by 25.56%and 10.00%in vivo experiment after 7 days respectively. The strain UV20-13 was tested by the experiments of subculture and dynamic growth,and the results showed that the strain UV20-13 was better than K.apiculata in the growth characteristics,and it did not appear retrogression,reversion mutation ect after subculturing 10 generations.Therefore the strain UV20-13 had genetic stability.