共查询到20条相似文献,搜索用时 0 毫秒
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Eugène K. Konan Justin Y. Kouadio Albert Flori Tristan Durand-Gasselin Alain Rival 《In vitro cellular & developmental biology. Plant》2007,43(5):456-466
In vitro rooting of oil palm shoots derived from somatic embryos was achieved through a single-phase protocol in which three shoots
are cultured in the same culture tube on an α-naphtaleacetic acid-enriched culture medium. Rooting performance was dependent
on both the genetic origin and initial size of the shoot explants. All shoots from a given tube showed a tendency to give
roots of the same type, independent of the original size of the explant. Whatever the clonal line, longer-size shoots (L-type:
>9 cm) showed higher rooting rates than medium-size (M-type: 7–9 cm) and short-size ones (S-type: 5–7 cm). When groups of
three shoots from the same clonal line were rooted together in the same culture tube, the combination of plant size within
the group impacted overall quality of rooting. Within triplets of shoots containing more than one short individual, the probability
of obtaining adequate rooting was low. Similarly, when more than one long shoot was included in the triplet rooting, quality
was also poor. By avoiding such combinations, the rate of well-rooted plantlets increased by 25%, with a maximum of 66% when
triplets of S/M/L combination were used. Smaller shoots, which usually showed poor rooting performance, were therefore found
to benefit from the presence of their neighbors. This interaction between the sizes of individuals in a given tube was found
to be associated with a within-tube correlation effect, a phenomenon previously described as “event coupling,” which was estimated
using a distorted binomial-type distribution of probabilities. The resulting calculation of a coupling factor (average r = 0.60) explains the behavior of shoots within the same culture tube and their average rooting performance. Modeling of the
interactions that occurred during in vitro rooting is described here and is recommended for improvement of this critical step in micropropagation. 相似文献
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Microcystis viridis lectin (MVL), a sugar-binding protein originally isolated from freshwater blue-green algae Microcystis viridis, has been reported to have potent anti-HIV activity. In this paper, we described the expression and purification of recombinant-MVL
(R-MVL) gene in E. coli. The results demonstrated that the R-MVL in shake flask cultures was primarily expressed either in the form of inclusion
bodies at 37°C or in the soluble fraction at 23 °C. Secondly, a one-step purification based on nickel-affinity chromatography
was employed and 15 mg of highly purified (>95%) R-MVL from 1 l of cell cultures was yielded. The purified R-MVL was then
subjected to MALDI-TOF–MS analysis for protein identification. In conclusion, for the first time, the R-MVL was successfully
cloned and expressed in E. coli, which is useful for further study and large-scale cost-effective production of MVL protein. 相似文献
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V. N. Verbenko L. V. Kuznetsova E. P. Krupyan V. I. Shalguev 《Russian Journal of Genetics》2009,45(10):1192-1199
Plasmids pKS5 and pKSrec30 carrying normal and mutant alleles of the Deinococcus recA gene controlled by the lactose promoter slightly increase radioresistance of Escherichia coli cells with mutations in genes recA and ssb. The RecA protein of D. radiodurans is expressed in E. coli cells, and its synthesis can be supplementary induced. The radioprotective effect of the xenologic protein does not exceed
1.5 fold and yields essentially to the contribution of plasmid pUC19-recA1.1 harboring the E. coli recA
+ gene in the recovery of resistance of the ΔrecA deletion mutant. These data suggest that the expression of D. radiodurans recA gene in E. coli cells does not complement mutations at gene recA in the chromosome possibly due to structural and functional peculiarities of the D. radiodurans RecA protein. 相似文献
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A. B. Nur Azura M. Yusoff G. Y. A. Tan R. Jegadeesh D. R. Appleton S. Vikineswary 《Journal of industrial microbiology & biotechnology》2016,43(4):485-493
Actinomycete strain AUM 00500 was 99.5 % similar to Streptomyces sanglieri NBRC 100784T and was evaluated for antagonistic activity towards Ganoderma boninense, the causative fungus of basal stem rot of oil palm. The strain showed strong antifungal activity towards G. boninense in in vitro and SEM analysis showed various modes of inhibition of the fungus. Ethyl acetate extracts of single culture and inhibition zone of cross-plug culture by HPLC indicated that strain AUM 00500 produced two different antibiotics of the glutarimide group namely cycloheximide and actiphenol. In greenhouse trials, oil palm seed treated with spores of S. sanglieri strain AUM 00500 at 109 cfu/ml showed significant (P < 0.05) increase in oil palm seedlings growth when compared to the control. Streptomyces sanglieri strain AUM 00500 successfully colonised the epidermal surface of the roots of treated oil palm seedlings and it was recovered from root fragments plated on starch casein agar. 相似文献
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Treponema denticola is a small anaerobic spirochete often isolated from periodontal lesions and closely associated with periodontal diseases. This bacterium possesses a particular arginine peptidase activity (previously called BANA-peptidase or trypsin-like enzyme) that is common to the three cultivable bacterial species most highly associated with severe periodontal disease. We recently reported the identification of the opdB locus that encodes the BANA-peptidase activity of T. denticola through DNA sequencing and mutagenesis studies. In the present study, we report expression of T. denticola OpdB peptidase in Escherichia coli. The opdB PCR product was cloned into pET30b and then transformed into the E. coli BL21 (DE3)/pLysS expression strain. Assays of enzymatic activities in E. coli containing T. denticola opdB showed BANA-peptidase activity similar to that of T. denticola. Availability of this recombinant expression system producing active peptidase will facilitate characterization of the potential role of this peptidase in periodontal disease etiology. 相似文献
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Previous work from our laboratory has shown that most of Bacillus thuringiensis strains possess the ability to produce melanin in the presence of l-tyrosine at elevated temperatures (42 °C). Furthermore, it was shown that the melanin produced by B. thuringiensis was synthesized by the action of tyrosinase, which catalyzed the conversion of l-tyrosine, via l-DOPA, to melanin. In this study, the tyrosinase-encoding gene (mel) from B. thuringiensis 4D11 was cloned using PCR techniques and expressed in Escherichia coli DH5 . A DNA fragment with 1179 bp which contained the intact mel gene in the recombinant plasmid pGEM1179 imparted the ability to synthesize melanin to the E. coli recipient strain. The nucleotide sequence of this DNA fragment revealed an open reading frame of 744 bp, encoding a protein of 248 amino acids. The novel mel gene from B.thuringiensis expressed in E. coli DH5 conferred UV protection on the recipient strain. 相似文献
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Lee JH Moon YH Kim N Kim YM Kang HK Jung JY Abada E Kang SS Kim D 《Biotechnology letters》2008,30(4):749-754
The gene encoding sucrose phosphorylase (742sp) in Leuconostoc mesenteroides NRRL B-742 was cloned and expressed in Escherichia coli. The nucleotide sequence of the transformed 742sp comprised an ORF of 1,458 bp giving a protein with calculated molecular mass of 55.3 kDa. 742SPase contains a C-terminal amino acid sequence that is significantly different from those of other Leu. mesenteroides SPases. The purified 742SPase had a specific activity of 1.8 U/mg with a K
m of 3 mM with sucrose as a substrate; optimum activity was at 37°C and pH 6.7. The purified 742SPase transferred the glucosyl
moiety of sucrose to cytosine monophosphate (CMP).
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
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Inui M Suda M Kimura S Yasuda K Suzuki H Toda H Yamamoto S Okino S Suzuki N Yukawa H 《Applied microbiology and biotechnology》2008,77(6):1305-1316
A recombinant butanol pathway composed of Clostridium acetobutylicum ATCC 824 genes, thiL, hbd, crt, bcd-etfB-etfA, and adhe1 (or adhe) coding for acetyl-CoA acetyltransferase (THL), β-hydroxybutyryl-CoA dehydrogenase (HBD), 3-hydroxybutyryl-CoA dehydratase
(CRT), butyryl-CoA dehydrogenase (BCD), butyraldehyde dehydrogenase (BYDH), and butanol dehydrogenase (BDH), under the tac promoter control was constructed and was introduced into Escherichia coli. The functional expression of these six enzymes was proved by demonstrating the corresponding enzyme activities using spectrophotometric,
high performance liquid chromatography and gas chromatography analyses. The BCD activity, which was not detected in E. coli previously, was shown in the present study by performing the procedure from cell extract preparation to activity measurement
under anaerobic condition. Moreover, the etfA and etfB co-expression was found to be essential for the BCD activity. In the case of BYDH activity, the adhe gene product was shown to have higher specificity towards butyryl-CoA compared to the adhe1 product. Butanol production from glucose was achieved by the highly concentrated cells of the butanologenic E. coli strains, BUT1 with adhe1 and BUT2 with adhe, under anaerobic condition, and the BUT1 and BUT2 strains were shown to produce 4 and 16-mM butanol with 6- and 1-mM butyrate
as a byproduct, respectively. This study reports the novel butanol production by an aerobically pregrown microorganism possessing
the genes of a strict anaerobe, Clostridium acetobutylicum. 相似文献
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Functional expression of lipase from Burkholderia sp. C20 (Lip) in various cellular compartments of Escherichia coli was explored. The poor expression in the cytoplasm of E. coli was improved by several strategies, including coexpression of the cytoplasmic chaperone GroEL/ES, using a mutant E. coli host strain with an oxidative cytoplasm, and protein fusion technology. Fusing Lip with the N-terminal peptide tags of T7PK,
DsbA, and DsbC was effective in enhancing the solubility and biological activity. Non-fused Lip or Lip fusions heterologously
expressed in the periplasm of E. coli formed insoluble aggregates with a minimum activity. Biologically active and intact Lip was obtained upon the secretion into
the extracellular medium using the native signal peptide and the expression performance was further improved by coexpression
of the periplasmic chaperon Skp. The extracellular expression was even more effective when Lip was secreted as a Lip–HlyA
fusion via the α-hemolysin transporter. Finally, Lip could be functionally displayed on the E. coli cell surface when fused with the carrier EstA. 相似文献
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O. V. Sergeeva D. O. Bredikhin M. V. Nesterchuk M. V. Serebryakova P. V. Sergiev O. A. Dontsova 《Biochemistry. Biokhimii?a》2018,83(3):270-280
Proteins containing the NIF3 domain are highly conserved and are found in bacteria, eukaryotes, and archaea. YbgI is an Escherichia coli protein whose gene is conserved among bacteria. The structure of YbgI is known; however, the function of this protein in cells remains obscure. Our studies of E. coli cells with deleted ybgI gene suggest that YbgI is involved in formation of the bacterial cell wall. 相似文献
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Preeyanan Anwised Nisachon Jangpromma Theeranan Temsiripong Rina Patramanon Sakda Daduang Sarawut Jitrapakdee Tomohiro Araki Sompong Klaynongsruang 《The protein journal》2016,35(4):256-268
Recombinant Crocodylus siamensis hemoglobin (cHb) has been constructed and expressed using Escherichia coli as the expression system in conjunction with a trigger factor from the Cold-shock system as the fusion protein. While successful processing as soluble protein in E. coli was achieved, the net yields of active protein from downstream purification processes remained still unsatisfactory. In this study, cHb was constructed and expressed in the eukaryotic expression system Pichia pastoris. The results showed that cHb was excreted from P. pastoris as a soluble protein after 72 h at 25 °C. The amino acid sequence of recombinant cHb was confirmed using LC–MS/MS. Indeed, the characteristic of Hb was investigated by external heme incorporation. The UV–Vis profile showed a specific pattern of the absorption at 415 nm, indicating the recombinant cHb was formed complex with heme, resulting in active oxyhemoglobin (OxyHb). This result suggests that the heme molecules were fully combined with heme binding site of the recombinant cHb, thus producing characteristic red color for the OxyHb at 540 and 580 nm. The results revealed that the recombinant cHb was prosperously produced in P. pastoris and exhibited a property as protein–ligand binding. Thus, our work described herein offers a great potential to be applied for further studies of heme-containing protein expression. It represents further pleasing option for protein production and purification on a large scale, which is important for determination and characterization of the authenticity features of cHb proteins. 相似文献
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A novel antifreeze protein cDNA was cloned by RT-PCR from the larva of the yellow mealworm Tenebrio molitor. The coding fragment of 339 bp encodes a protein of 112 amino acid residues and was fused to the expression vectors pET32a
and pTWIN1. The resulted expression plasmids were transformed into Escherischia coli strains BL21 (DE3), ER2566, and Origami B (DE3), respectively. Several strategies were used for expression of the highly
disulfide-bonded β-helix-contained protein with the activity of antifreeze in different expression systems. A protocol for
production of refolded and active T. molitor antifreeze protein in bacteria was obtained. 相似文献
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The gas-exchange characteristics, leaf water potential and chlorophyll (Chl) a fluorescence of oil palm (Elaeis guineensis Jacq.) seedlings subjected to water stress and recovery were investigated in a greenhouse experiment. At 24 days after imposition
of stress, leaf water potential in water-stressed seedlings was doubled compared to that of control and there was a drastic
decline in gas-exchange parameters viz. photosynthesis, transpiration, and stomatal conductance. Water stress did not irreversibly affect gas-exchange parameters
and quantum efficiency of photosystem II, as seedlings exhibited total recovery of photosynthetic apparatus by 12th day of rehydration. These findings indicate that oil palm exhibits physiological plasticity to water stress during the seedling
stage. 相似文献
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An effective protein expression system was constructed in Escherichia coli using the promoter of the tyrosine phenol-lyase (tpl) gene of Erwinia herbicola. This system involves a mutant form of the TyrR protein with an enhanced ability to activate tpl and the TutB protein with an ability to transport L-tyrosine (an inducer of Tpl). The highest expression level obtained for this system was more than twice that obtained for
the tac system, although it was lower than the level obtained for the T7 system, as revealed with the lac-reporter assay and SDS-polyacrylamide gel electrophoresis. 相似文献
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Si-Xiu Liu Zhong-Ping Fu Rui-Min Mu Zhi-Bi Hu Fu-Jun Wang Xiang-Rong Wang 《Molecular biology reports》2010,37(4):1781-1786
A nucleic acid sequence MC, encoding Momordica Chanrantia anti-hyperglycaemic peptide MC6 (accession: AAX06814) synthesized according to Escherichia coli preferred codons, was cloned and expressed in E. coli. Recombinant protein pQE8-MC (about 3.5 kDa) was purified and analyzed by 20% SDS–PAGE and western blot. It revealed that
the expressed pQE8-MC had good solubility in aqueous media. An HPLC assay was used to confirm the expression of pQE8-MC. Subsequent
pharmacological activity assay revealed a significant hypoglycemic effect of low dose treatments of pQE8-MC on male kunming
mice. Four hours after an intravenous tail injection, the blood sugar levels of mice treated with pQE8-MC saline solution
A3 (1 mg/kg BW) decreased greatly (P < 0.01) relative to the levels of a control group. This suggests that pQE8-MC, expressed in bioengineered E. coli, has a similar hypoglycemic function to the natural protein MC6 from M. Chanrantia. These results reveal the possibility of using bio-engineered bacteria as an anti-diabetic agent. 相似文献
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Extracellular protein profiles were compared for broth-grown cultures of Burkholderia pseudomallei and its avirulent close relative Burkholderia thailandensis. A number of protein bands were present in the B. pseudomallei profile but absent or less abundant in the B. thailandensis profile. Four such prominent secreted proteins were identified by using N-terminal sequencing coupled to searches of the B. pseudomallei genome sequence database. The genes for two proteins with similarity to carbohydrate-binding proteins, and a further protein homologous to known bacterial collagenases, were present in both B. pseudomallei and B. thailandensis. The putative collagenase gene was cloned and expressed as a fusion protein in Escherichia coli. Cell lysates from Escherichia coli containing the recombinant protein exhibited detectable gelatinase and collagenase activities. 相似文献