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1.
Gene flow from transgenic plants to compatible wild relatives is one of the major impediments to the development of the culture
of genetically engineered crop plants. In this work, the flow of EPSPS (conferring resistance to glyphosate) gene of transgene Brassica napus toward the untransgene B. napus and wild relative species Orychophragmus violaceus in an open field (1 ha) was studied. The data related to only the 2004 and 2005 autumn season on one location of southwest
of China. Pollen dispersal and fertilization of the target plants were favored and a detailed analysis of the hybrid offspring
was performed. In field, the data studied show that the gene flow frequency was 0.16% between GM and non-GM B. napus at a distance of 1 m from the transgenic donor area. The crosspollination frequency was 0.05% between GM and non-GM B. napus at a distance of 5 m from the transgenic donor area. At a distance of 10 m, no crosspollination was observed. According to
the results of this study, B. napus transgene flow was low. However, the wild relative species O. violaceus could not be fertilized by the transgenic pollen of B. napus, no matter what the distance was. 相似文献
2.
Toyota A Akiyama H Sugimura M Watanabe T Kikuchi H Kanamori H Hino A Esaka M Maitani T 《Bioscience, biotechnology, and biochemistry》2006,70(4):821-827
Because the labeling of grains and feed- and foodstuffs is mandatory if the genetically modified organism (GMO) content exceeds a certain level of approved genetically modified varieties in many countries, there is a need for a rapid and useful method of GMO quantification in food samples. In this study, a rapid detection system was developed for Roundup Ready Soybean (RRS) quantification using a combination of a capillary-type real-time PCR system, a LightCycler real-time PCR system, and plasmid DNA as the reference standard. In addition, we showed for the first time that the plasmid and genomic DNA should be similar in the established detection system because the PCR efficiencies of using plasmid DNA and using genomic DNA were not significantly different. The conversion factor (Cf) to calculate RRS content (%) was further determined from the average value analyzed in three laboratories. The accuracy and reproducibility of this system for RRS quantification at a level of 5.0% were within a range from 4.46 to 5.07% for RRS content and within a range from 2.0% to 7.0% for the relative standard deviation (RSD) value, respectively. This system rapidly monitored the labeling system and had allowable levels of accuracy and precision. 相似文献
3.
Transgenic cotton expressing Cry10Aa toxin confers high resistance to the cotton boll weevil
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Thuanne Pires Ribeiro Fabricio Barbosa Monteiro Arraes Isabela Tristan Lourenço‐Tessutti Marilia Santos Silva Maria Eugênia Lisei‐de‐Sá Wagner Alexandre Lucena Leonardo Lima Pepino Macedo Janaina Nascimento Lima Regina Maria Santos Amorim Sinara Artico Márcio Alves‐Ferreira Maria Cristina Mattar Silva Maria Fatima Grossi‐de‐Sa 《Plant biotechnology journal》2017,15(8):997-1009
4.
Chang-Gi Kim Dae In Kim Hyo-Jeong Kim Ji Eun Park Bumkyu Lee Kee Woong Park Soon-Chun Jeong Kyung Hwa Choi Joo Hee An Kang-Hyun Cho Young Soon Kim Hwan Mook Kim 《Journal of Plant Biology》2009,52(3):251-258
We conducted a 2-year field assessment of the gene flow from genetically modified (GM) chili pepper (Capsicum annuum L.), containing the PepEST (pepper esterase) gene, to a non-GM control line “WT512” and two commercial hybrid cultivars, “Manidda” and “Cheongpung Myeongwol
(CM).” After seeds were collected from the pollen-recipient non-GM plants, hybrids between them and the GM peppers were screened
by a hygromycin assay. PCR with the targeting hpt gene was performed to confirm the presence of transgenes in hygromycin-resistant seedlings. Out of 7,071 “WT512” seeds and
6,854 “Manidda” seeds collected in 2006, eight and 12 hybrids, respectively, were detected. In 2007, 33 hybrids from 3,456
“WT512” seeds and 50 hybrids from 3,457 “CM” seeds were found. The highest frequency of gene flow, 6.19%, was observed in
that 2007 trial. These results suggest that a limited isolation distance would be sufficient to prevent gene flow from GM
to conventionally bred chili peppers. 相似文献
5.
Su-Il Do 《Glycoconjugate journal》2009,26(5):567-575
Novel chimeric lacdiNAc (GalNAc(β1-4)GlcNAc) synthase (c-LacdiNAcS) was generated by gene fusion of α-lactalbumin (α-LA) and
β1,4-galactosyltransferase 1 (β1,4-GalT1). c-LacdiNAcS was expressed in Lec8 Chinese hamster ovary (Lec8 CHO) cells and exhibited
N-acetylgalactosaminyltransferase (GalNAcT) activity in the absence of exogenous α-LA as well as other glycosyltransferase
activities including lactose synthase (LacS), and β1,4-GalT. These glycosyltransferase activities of c-LacdiNAcS were compared
to those activities induced in LacS system under the co-presence of bovine β1,4-GalT1 and α-LA, indicating that each domain
of α-LA and β1,4-GalT1 on c-LacdiNAcS is not only folding correctly, but also interacting together. Furthermore, c-LacdiNAcS
was found to be auto-lacdiNAcylated and can synthesize lacdiNAc structures on cellular glycoproteins, demonstrating that GalNAcT
activity of c-LacdiNAcS is functional in Lec8 CHO cells. 相似文献
6.
R. B. Turley 《Biologia Plantarum》2008,52(4):759-762
Two dimensional polyacrylamide gel electrophoresis (2D-PAGE) was used to identify differentially expressed proteins in wild-type
(DP 5690) and fiberless (SL 1-7-1) cotton ovules. One protein, designated V2 was unique to ovules of the fiber producing DP
5690 line. The protein was purified from 2D-PAGE of 4 d post anthesis DP 5690 ovules and partially sequenced. The short amino
acid sequence was nearly identical to the deduced amino acid sequence for cotton phenylcoumaran benzylic ether reductase (PCBER)
protein. A consensus sequence was assembled from ESTs encoding cotton PCBER genes, primers were designed, and a full length
gene was amplified from plasmid DNA from a 72 h etiolated cotton cotyledon library. The polymerase chain reaction generated
a 950 bp product with unique EcoRI (5′) and (3′) KpnI restriction sites for directional insertion into the expression vector pPICZA. Nucleotide sequencing was performed, and
the full length coding region was 924 bp encoding a protein of 308 amino acids. The molecular mass and pI measured (2D PAGE)
were similar to the theoretical protein. 相似文献
7.
This study was conducted to compare the safety of soybean meal prepared from genetically modified (GM) glyphosate-tolerant (Roundup Ready; RR) soybeans and conventional soybeans. Eighty Sprague-Dawley rats (40 males and 40 females) were randomly allotted to one of four groups according to sex and body weight for a 13-week feeding experiment. The rats were fed corn-based diets containing 60% conventional soybean meal, a mixture of 30% conventional and 30% RR soybean meal, 60% or 90% RR soybean meal. All diets were adjusted to an identical nutrient level except the 90% RR diet. The two soybean meals were similar in chemical analysis and amino acid composition. During the 13-week growth trial, body weight (P?0.05) and feed intake (P?0.05) decreased only in rats fed with 90% RR soybean meal at the first week. No treatment-related deaths occurred during the experiment. Gross necropsy findings, haematological or urinalysis values and clinical serum parameters showed no meaningful differences between rats fed the control and RR soybean meals. A 145?bp of cp4 epsps gene specific for the GM constructs from RR soybean meal or a 407?bp of lec gene from endogenous soybean DNA could not be detected in investigated masseter muscle samples. No adverse effects of glyphosate-tolerant soybean meal on rats were seen even at levels as high as 90% of the diet. 相似文献
8.
Yan Xunyou Zhao Hongliang Zhang Weiguang Xue Chong Liu Zhimin 《Frontiers of Biology in China》2007,2(2):170-175
To obtain human tissue inhibitor of metalloproteinase-2 (TIMP-2) cDNA and the secretory expression of TIMP-2 gene in Pichia pastoris, we designed and synthesized a 618 base pairs artificial gene coding for the TIMP-2 with a computer-aided design method using
a standard chemical synthesis technique, which was composed of frequently used codons in the highly expressed Pichia pastoris genes. Then the synthetic gene encoding TIMP-2 was checked by means of dideoxynucleotide sequencing. The verified gene of
TIMP-2 was cloned to the Escherichia coli-yeast shuttle vector of pPIC9 to construct a recombinant plasmid pPIC9-T2. The plasmid was transformed into GS115 cells of
the methylotrophic yeast, Pichia pastoris by electroporation, and we got the expression cell through phenotype selection and induction with methanol. Separation, purification,
and bioactivity analysis of the expressed products were performed.
__________
Translated from Microbiology, 2006, 33(1): 1–6 [译自: 微生物学通报] 相似文献
9.
10.
Ortiz-Alvarado R Gonzalez-Hernandez GA Torres-Guzman JC Gutierrez-Corona JF 《Current microbiology》2006,52(3):178-181
Mucor circinelloides transformants prototrophic to leucine and resistant to carboxine (Leu+ Cbxr) have been obtained by treatment of protoplasts with plasmid constructs containing homologous leuA gene and adjacent autonomously replicating sequences (ARS) element combined with the Cbxr(carboxine-resistance) gene of Ustilago maydis and ARS sequences from this basidiomycete (plasmid pGG37) or from the 2 μ plasmid of Saccharomyces cerevisiae (plasmid pGG43). The presence in the same plasmid molecule of the M. circinelloides leuA gene and adjacent ARS element together with heterologous ARS elements produced an increase in the transformation frequency
of about 65–120%. The presence of autoreplicating plasmid molecules in the transformants was demonstrated by mitotic stability
experiments, by Southern analysis, and by the rescue of plasmids from transformed bacterial cells. 相似文献
11.
Chang-Gi Kim Bumkyu Lee Dae In Kim Ji Eun Park Hyo-Jeong Kim Kee Woong Park Hoonbok Yi Soon-Chun Jeong Won Kee Yoon Chee Hark Harn Hwan Mook Kim 《Journal of Plant Biology》2008,51(1):74-77
Gene flow from genetically modified (GM) crops to conventional non-GM crops is a serious concern for protection of conventional
and organic farming. Gene flow from GM watermelon developed for rootstock use, containing cucumber green mottle mosaic virus
(CGMMV)-coat protein (CP) gene, to a non-GM isogenic control variety “Clhalteok” and grafted watermelon “Keumcheon” was investigated
in a small scale field trial as a pilot study. Hybrids between GM and non-GM watermelons were screened from 1304 “Chalteok”
seeds and 856 “Keumcheon” seeds using the duplex PCR method targeting theCGMMV- CP gene as a marker. Hybrids were found in all pollen recipient plots. The gene flow frequencies were greater for “Chaiteok”
than for “KeumcheonD; with 75% outcrossing in the “Chaiteok” plot at the closest distance (0.8 m) to the GM plot. A much larger
scale field trial is necessary to identify the isolation distance between GM and non-GM watermelon, as the behaviors of insect
pollinators needs to be clarified in Korea. 相似文献
12.
The rpoS gene from Pseudomonas sp. M18, which encodes predicted protein (an alternative sigma factor s, σS, or σ38) with 99.5% sequence identity with RpoS from Pseudomonas aeruginosa PAO1, was first cloned. In order to investigate the mechanism of rpoS expression, an rpoS null mutant, named M18S, was constructed with insertion of aacC1 cassette bearing a gentamycin resistance gene. With introduction of a plasmid containing an rpoS′–′lacZ translational fusion (pMERS) to wild-type strain M18 or M18S, it was first found that β-galactosidase activity expressed
in strain M18S (pMERS) decreased to fourfold of that expressed in the strain M18 (pMERS). When strain M18S (pMERS) was introduced
with another plasmid pBBS containing the wild-type rpoS gene, its β-galactosidase expression level was enhanced and almost restored to that in strain M18 (pMERS). Similarly, expression
of β-galactosidase from a chromosomal fusion of the promoter of the wild-type rpoS gene with lacZ (rpoS–lacZ) was enhanced fivefold in the presence of a plasmid with the wild-type rpoS gene. With these findings, it is suggested that RpoS sigma factor may be involved in autoinducing its own gene expression
in Pseudomonas sp. M18. 相似文献
13.
Todd A. Gaines Alice A. Wright William T. Molin Lothar Lorentz Chance W. Riggins Patrick J. Tranel Roland Beffa Philip Westra Stephen B. Powles 《PloS one》2013,8(6)
Weed populations can have high genetic plasticity and rapid responses to environmental selection pressures. For example, 100-fold amplification of the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene evolved in the weed species Amaranthus palmeri to confer resistance to glyphosate, the world’s most important herbicide. However, the gene amplification mechanism is unknown. We sequenced the EPSPS gene and genomic regions flanking EPSPS loci in A. palmeri, and searched for mobile genetic elements or repetitive sequences. The EPSPS gene was 10,229 bp, containing 8 exons and 7 introns. The gene amplification likely proceeded through a DNA-mediated mechanism, as introns exist in the amplified gene copies and the entire amplified sequence is at least 30 kb in length. Our data support the presence of two EPSPS loci in susceptible (S) A. palmeri, and that only one of these was amplified in glyphosate-resistant (R) A. palmeri. The EPSPS gene amplification event likely occurred recently, as no sequence polymorphisms were found within introns of amplified EPSPS copies from R individuals. Sequences with homology to miniature inverted-repeat transposable elements (MITEs) were identified next to EPSPS gene copies only in R individuals. Additionally, a putative Activator (Ac) transposase and a repetitive sequence region were associated with amplified EPSPS genes. The mechanism controlling this DNA-mediated amplification remains unknown. Further investigation is necessary to determine if the gene amplification may have proceeded via DNA transposon-mediated replication, and/or unequal recombination between different genomic regions resulting in replication of the EPSPS gene. 相似文献
14.
D K-H Wong K H E Lam C K P Chan Y C V Wong W K R Wong J Hackett 《Journal of industrial microbiology & biotechnology》1998,21(1-2):31-36
A plasmid stabilization system, active in high copy-number plasmids, was cloned from the large resident plasmid, pSLT, of
Salmonella typhimurium. The ytl2 gene, together with a 249-bp region (termed incR) downstream of the gene, imparted >104-fold stability to a pBR322-based plasmid. The ytl2-incR region was then used to stabilize a recombinant plasmid carrying the human epidermal growth factor gene (with the Escherichia coli K-12 ompA signal sequence), behind the lacUV5 promoter. In shake flask tests to optimize expression of human epidermal growth factor, loss of recombinant plasmid was <1%
when growth (both before and after induction with isopropyl-β-d-galactopyranoside) took place even in the absence of antibiotic
selection, and the specific activity of secreted human epidermal growth factor was ca 20 μg per 108 cells at harvest, compared to a figure of ca 3 μg per 108 cells when a comparable plasmid, but devoid of the ytl2-incR region, was employed, as outgrowth of plasmid-free cells after induction severely compromised the specific activity of the
secreted product.
Received 2 June 1998/ Accepted in revised form 15 July 1998 相似文献
15.
Isabel M.G. Padilla Agnieszka Golis Adele Gentile Carmine Damiano Ralph Scorza 《Plant Cell, Tissue and Organ Culture》2006,84(3):309-314
To determine the optimum conditions for Agrobacterium-mediated gene transfer, peach explants including cotyledons, embryonic axes and hypocotyl slices from non-germinated seeds
and epicotyl internode slices from germinating seeds were exposed to Agrobacterium-mediated transformation treatments. The GUS (uidA) marker gene was tested using two different A. tumefaciens strains, three plasmids and four promoters [CaMV35s, (Aocs)3AmasPmas (“super-promoter”), mas-CaMV35s, and CAB]. GFP was tested with six A.␣tumefaciens strains, one plasmid (pLC101) and the doubleCaMV35s (dCaMV35s) promoter. The CaMV35s promoter produced more GUS expression
than the CAB promoter. A. tumefaciens strains EHA105 and LBA4404 harboring the same plasmid (pBIN19) differed in their effects on GUS expression suggesting an
interaction between A. tumefaciens strain and plasmid. A combination of A. tumefaciens EHA105, plasmid pBIN19 and the CaMV35s promoter produced the highest rates of transformation in peach epicotyl internodes
(56.8%), cotyledons (52.7%), leaves (20%), and embryonic axes (46.7%) as evaluated by the percentage of explants expressing
GUS 14 days after co-cultivation. GFP expression under the control of the dCaMV35s promoter was highest for internode explants
but only reached levels of 18–19%. When GFP-containing plasmid pCL101 was combined with each of five A. tumefaciens strains the highest levels of transformation were 20–21% (internode and cotyledons, respectively). When nine peach genotypes
were co-cultivated with A. tumefaciens strain EHA105 and GFP-containing plasmid pCL101 the highest levels of transformation were 26–28% (cotyledons and internodes,
respectively). While GFP represents a potentially useful transformation marker that allows the non-destructive evaluation
of transformation, rates of GFP transformation under the conditions of this study were low. It will be necessary to optimize
expression of this marker gene in peach. 相似文献
16.
The gene which encodes (R)-specific carbonyl reductase (rCR) from Candida parapsilosis CCTCC M203011 was cloned, sequenced and compared with genes from the GenBank. The results indicated that rCR gene was 1011
bp, encoding a protein of 336 amino acids with a molecular weight of 35.9 kDa, and its nucleotide sequence showed 99% similarity
to those of other members of the alcohol dehydrogenase superfamily. The rCR gene could express in recombinant strain Escherichia coli JM109, and the expression plasmid could produce (R)-1-pheny-1,2-ethanediol (100% e.e., 80.14% yield) from β-hydroxyacetophenone without any additive to regenerate NAD+ from NADH.
__________
Translated from Microbiology, 2006, 33(4): 112–118 [译自: 微生物学通报] 相似文献
17.
Species of the Bacteroides fragilis group are considered the most common anaerobe in human and animal infections and also harbor plasmids conferring resistance
to several antibiotics. In this study, resistance to cefoxitin, plasmid profile and β-lactamase production in species of the
B. fragilis group isolated from intestinal tracts of calves were evaluated. One hundred sixty-one B. fragilis group bacteria isolated from calves with and without diarrhea were analyzed. Cefoxitin susceptibility was performed using
an agar dilution method, β-lactamase production by using a nitrocefin method, and plasmid extraction by using a commercial
kit. Minimal inhibitory concentration values for cefoxitin ranged from 32 to > 512 μg/ml, and 47 bacteria (29.2%) were resistant
to cefoxitin (breakpoint 16 μl). Only seven isolates harbored plasmids varying from 6.0 to 5.0 kb, and a 5.5-kb plasmid in
B. vulgatus Bd26e and B. fragilis Bc5j might be related to cefoxitin resistance. β-lactamase was detected in 33 (70.2%) isolates. The cepA gene was observed in total DNA and in the 5.5-kb plasmid. The plasmid presence in organisms isolated from cattle may be important
in ecologic terms, and it needs further study. 相似文献
18.
Qualitative and Quantitative PCR Methods for Event-specific Detection of Genetically Modified Cotton Mon1445 and Mon531 总被引:12,自引:0,他引:12
Based on the DNA sequences of the junctions between recombinant and cotton genomic DNA of the two genetically modified (GM)
cotton varieties, herbicide-tolerance Mon1445 and insect-resistant Mon531, event-specific primers and probes for qualitative
and quantitative PCR detection for both GM cotton varieties were designed, and corresponding detection methods were developed.
In qualitative PCR detection, the simplex and multiplex PCR detection systems were established and employed to identify Mon1445
and Mon531 from other GM cottons and crops. The limits of detection (LODs) of the simplex PCR were 0.05% for both Mon1445
and Mon531 using 100 ng DNA templates in one reaction, and the LOD of multiplex PCR analysis was 0.1%. For further quantitative
detection using TaqMan real-time PCR systems for Mon1445 and Mon531, one plasmid pMD-ECS, used as reference molecule was constructed,
which contained the quantitative amplified fragments of Mon1445, Mon531, and cotton endogenous reference gene. The limits
of quantification (LOQs) of Mon1445 and Mon531 event-specific PCR systems using plasmid pMD-ECS as reference molecule were
10 copies, and the quantification range was from 0.03 to 100% in 100 ng of the DNA template for one reaction. Thereafter,
five mixed cotton samples containing 0, 0.5, 0.9, 3 and 5% Mon1445 or Mon531 were quantified using established real-time PCR
systems to evaluate the accuracy and precision of the developed real-time PCR detection systems. The accuracy expressed as
bias varied from 1.33 to 8.89% for tested Mon1445 cotton samples, and from 2.67 to 6.80% for Mon531. The precision expressed
as relative standard deviations (RSD) were different from 1.13 to 30.00% for Mon1445 cotton, and from 1.27 to 24.68% for Mon531.
The range of RSD was similar to other laboratory results (25%). Concluded from above results, we believed that the established
event-specific qualitative and quantitative PCR systems for Mon1445 and Mon531 in this study are acceptable and suitable for
GM cotton identification and quantification. 相似文献
19.
This study focused on finding a novel sensitive method to determine the methylation modification at DNA dam (GATC) sites in Escherichia coli. A new plasmid which contained three GATC sites recognized by restriction enzyme BclI and one GAATTC site recognized by EcoRI
was transformed into E. coli stains AB1157(dam
+) and GM2929(dam
−) respectively. Then the plasmid DNA was digested by restriction enzyme BclI(T*GATCA), which was sensitive to methylation.
The results showed that the plasmid derived from AB1157 was not digested while that from GM2929 was, for the methylation level
of the former was high while the latter was low. So by detecting the methylation of plasmid transferred into the strain, we
could determine whether methylaion existed at DNA dam (GATC) site in E. coli. This method was effective and rapid; moreover, the digested fragments were not dispersive. It also made a basis for the
detection of whether methylation occurred in mode beings by low-energy ion beam.
The article is published in the original. 相似文献
20.
Jari Olavi Vehmaanperä Matti Pellervo Korhola 《Applied microbiology and biotechnology》1986,23(6):456-461
Summary Theα-amylase gene ofBacillus amyloliquefaciens has previously been cloned into pUB110 to give the recombinant plasmid, pKTH10 (Palva 1982. Gene 19:81–87). Strains transformed
by this plasmid are promising candidates for industrialα-amylase production. The stability of pKTH10 was determined in variousB. subtilis strains possessing specific alleles which affect the level ofα-amylase secretion.B. subtilis strains carrying pKTH10 were cultivated in starch-containing medium for up to 50 generations without antibiotic selection
and then screened for the presence of pKTH10. The plasmid proved stable enough (< 1.0% cured after 50 generations) for industrial
batchwise enzyme production in two strains, but in asacU9 strain (thesacU9 mutation increases concominantly the production ofα-amylase levansucrase and proteases) 99.9% of cells had lost pKTH10 after 50 generations, although the parental plasmid (pUB110)
was stable in this strain (0.09% cured after 50 generations). The instability of pKTH10 in thesacU9 strain seems somehow to be related to high expression of the clonedα-amylase gene: when grown in a medium restrictingα-amylase production, only 0.53% ofsacU9 cells had lost pKTH10 after 50 generations. 相似文献