CN102617711A - Cation diffusion assisting protein DbsCDF and encoding gene and application thereof - Google Patents
Cation diffusion assisting protein DbsCDF and encoding gene and application thereof Download PDFInfo
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Abstract
The invention discloses a metal cadmium resistance-relevant protein DbsCDF and an encoding gene and an application thereof. The amino acid sequence of the cadmium resistance-relevant protein DbsCDF is shown as SEQIDNO:1, and the encoding gene is shown as SEQIDNO:2. As proved by an escherichia coli experiment, the resistance of heavy metal cadmium can be enhanced through expression in escherichia coli. The DbsCDF encoding gene disclosed by the invention can be used for enhancing the resistance of microorganisms and plants to heavy metal cadmium, and is further used for clearing heavy metal pollution, and an excellent biological resource with high performance is provided for biological repair.
Description
Technical field
The invention belongs to microbiological genetic engineering and biological prosthetic field.Specifically; The invention provides and live in acidic mine waste water (acid mining drainage; AMD) a kind of gene to the heavy metal cadmium resistance that a kind of unknown Institute of Micro-biology is contained is the sequence of supporing yang ion diffusion protein gene (DbsCDF), and this sequence is inferred the aminoacid sequence of encode protein molecule.The present invention has great application value in raising mikrobe and plant aspect the resistance of heavy metal cadmium.
Background technology
Along with development and use, the industrial development of Mineral resources, the heavy metal pollution on the environment is on the rise, and heavy metal pollution of soil has become the problem of a harm global environment quality.Heavy metal-polluted soil can influence growth and development of plant, reduces output and the quality of farm crop, has brought serious economy loss.In addition, in plant materials, accumulated by the crop of heavy metal pollution of soil, and pass through the food chain enrichment in human body and animal body, harm human and livestock health, cause cancer and other diseases etc.Improvement heavy metal contamination is very urgent, during various recovery techniques and measure are being studied and used.National governments and scientist put forth effort to address this problem through two approach: one is to utilize physics, the heavy metal contamination that the method for chemistry attempts to remove soil or water body: two for utilizing modern biotechnology to remove pollution.Since the eighties in 20th century, processing costs is low, little to environmental influence, the efficient advantages of higher because of it has for bioremediation technology, more and more receives numerous scientific and technical personnel's extensive concern.Biological prosthetic generally is divided into phytoremediation, animal is repaired and mikrobe is repaired three types, and wherein the reparation of phytoremediation and mikrobe is the focus of research.It is exactly to utilize mikrobe with the contaminant degradation in the environment or be converted into the process of other innoxious substances that mikrobe is repaired.In recent years, based on the mechanism of action of mikrobe to heavy metal, the economic worth heavy metal is arranged is that the biologic treating technique of purpose reaches its maturity to repair poisonous and harmful metallic pollution or recovery.Phytoremediation refers to utilize plant to go to administer the technology of the pollution in the media such as water body, soil and bed mud.Yet the biology that is used for the heavy metal contamination reparation tends to receive the murder by poisoning of heavy metal, and poor growth, living weight are little, even can not survive, so the toxic action of heavy metal on plants and mikrobe is the key constraints of biological prosthetic.
Heavy metal is to study the The Molecular Biology Mechanism of tolerance heavy metal in the solution biological prosthetic to the fundamental way of the toxic action of biology; The clone obtains to be used for the transgenic engineering biology of biological prosthetic excellent property to the key gene of heavy metal tolerance through genetic engineering means.
Owing to technical reason, up to date, but the utilization of microbial gene resource mainly is confined to culturing micro-organisms.Yet, culturing micro-organisms only account for the occurring in nature mikrobe less than 1%, therefore the grand genome of mikrobe in the various habitats is a huge and genetic resources storehouse that do not excavate.Extreme environment has abundant Microbial resources, in the middle of many genes relevant with crucial vital process with adverse circumstance in secular adaptive evolution, obtained stronger patience potential, excavating these resistant genes has become international important research focus.Acidic mine waste water (AMD) is the important system of extreme habitat microbiological research.AMD derives from mining activity and makes and contain sulfur mineral and (be mainly pyrite, FeS
2) be exposed in the sky G&W, rapid oxidation is produced due to the acid under the microorganism catalysis effect, and its pH value and is rich in vitriol and heavy metals such as Pb, Zn, Cu, Cd and Ni generally about 1-4, is one of serious environmental problems of facing of mining industry.The prokaryotic micro-organisms of in AMD, surviving has formed some unique mechanism gradually in the evolution of long period of time process, compel to tackle multiple extreme environment such as low pH value, high salinity and high heavy metal association.Therefore, the AMD habitat becomes the adversity gene storehouse that has characteristic and enrich.
(cation diffusion facilitator, CDF) family has anti-heavy metal effect to supporing yang ion diffusion albumen, is present on a variety of biological fluid vacuolar membranes.Non-direct evidence shows: these protein have translocation, both can metals ion be transported in the cell, also can metals ion be exported cell.And CDF protein helps most organisms to Zn
2+, Cd
2+, Co
2+, Ni
2+Tolerance.The CDF family protein of different sources structurally has some general character: N-end signal sequence, stride the film district for 6, and in most of eukaryotes, also there is His enrichment territory in the born of the same parents in C-ion in Dragon Boat Festival domain output, and in prokaryotic organism, does not have.In Arabidopis thaliana, there is the protein of 8 genes encodings to belong to CDF family; 4 kinds have the said structure characteristic; And other several kinds (structure of AMtTPc1～AtMTPc4) is slightly different: comprise 4-5 membrane spaning domain, wherein 3 kinds only have part N-end signal sequence, and shortage His enrichment territory.The major function of Arabidopis thaliana AtMtTP1 is to improve the metal patience of cell.Have research to show: AtMtTP1 can be to Zn
2+Murder by poisoning play detoxification.If in smooth xenopus leavis oocytes, express AtMTP1, but the more Zn of enrichment then
2+In addition, through the expression of hook khuskhus cDNA library in cereuisiae fermentum will be arranged, identify 4 kinds of relevant CDF albumen, wherein ShMTP1 is to Mn
2+Patience is arranged.And people such as Persuns isolate the TgMTPs gene from wild mustard, and the CDF albumen of its coding can recover cereuisiae fermentum COT1 and ZRC1 (in the yeast to Zn
2+And Co
2+Indefatigable CDF albumen) ability of the anti-heavy metal of two mutants.
Therefore the CDF gene is played the part of very important role in the plant under heavy metal stress.Yet in mikrobe, what effect to be the heavy metal cadmium resistance played actually still unclear for the CDF gene up to now, also from the AMD mikrobe, is not cloned into the report of CDF gene.Is heavy metal content high among the AMD, but which gene pairs heavy metal resistance plays keying action in the AMD mikrobe? Does the CDF gene play an important role to AMD microbial growth and existence? These all are open questions still at present.
Summary of the invention
The object of the present invention is to provide the supporing yang ion diffusion albumen in the AMD mikrobe, and this proteic new gene of coding, basic substance established for developing gene engineering product from now on.
In one aspect of the invention, a kind of heavy metal cadmium resistance protein (supporing yang ion diffusion albumen in the acidic mine waste water mikrobe) is provided, has been called DbsCDF albumen in the present invention, it is a following protein molecular (i) or (ii):
(i) has described aminoacid sequence like SEQ ID NO:1;
(ii) going into the aminoacid sequence process replacement that SEQ ID NO:1 is limited, lack or superpose one or several amino acid derived protein and protein (i) have identical functions.
In another aspect of the present invention, a kind of dna molecular is provided, it comprises: the proteic nucleotide sequence of the described supporing yang ion diffusion of encoding.
Invention finds that DbsCDF albumen has the effect of cadmium metal resistance, can in administering Environmental Cadmium Pollution, use.
This proteic encoding sox has been protected in invention simultaneously, has the sequence shown in SEQ ID NO:2; And the application of this gene in administering Environmental Cadmium Pollution.
Invention, is found to coerce down at cadmium as subjects with the genetic engineering bacterium of expressing DbsCDF, and its cadmium tolerance is much higher than the non-genomic engineering bacteria.At concentration of cadmium ions is 200 μ M when following, when especially 150M is following, still has higher survival rate.
Compared with prior art, the present invention has following beneficial effect:
The present invention has cloned a new supporing yang ion diffusion protein gene from the AMD mikrobe, called after DbsCDF, and its resistance to heavy metal cadmium carried out functional analysis.Can improve in the intestinal bacteria resistance through this gene of intestinal bacteria transformation experiment proof to cadmium.
Said gene sequence or aminoacid sequence carry out transgenic exploitation gene engineering product mask has great application value; Can be used for cultivating the heavy metal super-enriched plant or the mikrobe of high-biomass specifically, be used for the restoration of the ecosystem of heavy-metal contaminated soil and water body.
Description of drawings
Phase when accompanying drawing 1 is the expression of DbsCDF in intestinal bacteria; Wherein:
M: molecular weight of albumen standard
1: BL21 (DE3) bacterial strain that carries empty carrier pET28a
2-9: BL21 (DE3) bacterial strain that carries pET28a-DbsCDF is distinguished abduction delivering 0,1,2,3,4,5,6,7 hour.
Accompanying drawing 2 is for expressing and the growth curve of intestinal bacteria under 150 μ M cadmiums are coerced of not expressing DbsCDF.
Accompanying drawing 3 is for expressing and the growing state of the intestinal bacteria of not expressing DbsCDF after cultivating 12 hours under the different cadmium concentrations.
Embodiment
The clone of embodiment 1DbsCDF gene complete sequence
Open-air microbiological specimens collection:
Lead/zinc ore is chosen the AMD of different souring stages (is standard with pH) in the Yunfu, uses the filter membrane of 0.22 μ m to collect the cell of 20LAMD the inside.In order to keep the complete preservation of nucleic acid, preserve sample and freeze in liquid nitrogen, take back the laboratory in 24 hours, and be put in-70 ℃ of refrigerator prolonged preservation.
The extraction of nucleic acid:
DNA extraction adopts the SET method, and process is following: in SET buffer, add N,O-Diacetylmuramidase and Proteinase K, behind the digestion 30min; 15, behind the centrifugal 15min of 000rpm with chloroform extracting 2 times, after spending the night with isopropanol precipitating; 75% ethanol cleans 2 times, is dissolved in the aqua sterilisa at last.Reclaim genomic dna with Qiagen tip-100 column purification, detect DNA quality and concentration with the nucleic acid-protein analyser.
Gene order-checking:
With press proof article in the GS FLX Titanium General Library Preparation Kit preparation of Roche company.Use Roche 454Genome Seqencer FLX sequenator,, obtain base sequence with Pyrobayer software through checking order.
Genome sequence is analyzed:
After removing low-quality sequencing result, genome is analyzed as follows:
Sequence assembly: use the GS De Novo Assembler Software of Euler-SR and 454 Corp. to carry out sequence assembly.Effect with the splicing of N50 index assessment;
Genome annotation: the whole genome sequence that will splice good mikrobe carries out genomic note with IMG and SEED system, finds new gene.
The clone of DbsCDF gene fragment:
Pass through PCR method: with the cloned plasmids that contains goal gene is template; Design a pair of primer (introducing different restriction enzyme sites respectively) by gene order at the upstream and downstream primer; PCR obtains the band of a 1kb size; And be cloned in PCR2.1 (available from the invitrogen company) carrier, sequence entrusts invitrogen company to measure.
The DbsCDF gene sequencing:
Sequencing result shows that DbsCDF gene size is 957bp (seeing SEQ ID NO:2), 318 amino acid (seeing SEQ ID NO:1) of encoding.According to psort analysis, the DbsCDF gene expression product is likely epicyte protein.
The functional analysis of embodiment 2DbsCDF gene
Utilize the intestinal bacteria transformation experiment to analyze the function of DbsCDF gene in the present embodiment.
Make up recombinant expression vector:
A pair of primer below the design is introduced the BamHI restriction enzyme site at the DbsCDF gene 5 ', and 3 ' introduces the XhoI restriction enzyme site.
DbsCDF-F(SEQ?ID?NO：3)：
5’CGCGGATCCATGGCACACGATTCTCATGACC3’
DbsCDF-R(SEQ?ID?NO：4)：
5’CCGCTCGAGTCAAACCCGATGATCCGGC3’
With the PCR2.1-DbsCDF carrier is that template is carried out PCR.Respectively PCR product and yeast shuttle expression carrier pET28a are carried out enzyme with BamHI and XhoI and cut, enzyme is cut product reclaim, connect and transformed into escherichia coli DH5 α.Cut evaluation through order-checking and enzyme, obtained the pET28a-DbsCDF recon.
Protein expression:
With recon pET28a-DbsCDF transformed into escherichia coli expression strain BL21 (DE3), through PCR checking screening positive clone.The thalline list spot that picking contains recombinant plasmid 37 ℃ of incubated overnight to the 10ml LB (containing Kan50 μ g/ml).1ml bacterium liquid joined contain 100ml LB substratum (containing Kan50 μ g/ml), 37 ℃ of concussions are cultured to OD600 and are about 0.4-1.0 (best 0.6, approximately need 2hr).
Adding IPTG was that 1mM induces to final concentration, whenever collected 1ml bacterium liquid at a distance from 1 hour, and centrifugal 12000g * 60s gathers in the crops deposition, and is resuspended with 80 μ l ddH2O, adds 20 μ l, 5 * SDS-PAGE sample-loading buffer, mixing, boiling water bath 10min.Get supernatant and do SDS-PAGE analysis (seeing accompanying drawing 1) as sample.
Transformant is to the mensuration of heavy metal cadmium resistance:
The thalline list spot that picking contains recombinant plasmid 37 ℃ of incubated overnight to the 10ml LB (containing Kan 50 μ g/ml).The 100ul bacterium is joined 10ml Cd
2+Concentration is respectively in the LB substratum (containing Kan 50 μ g/ml, IPTG1mM) of 0 μ M, 50 μ M, 100 μ M, 150 μ M, 200 μ M, and 37 ℃, 200rpm concussion were cultivated 12 hours, measured the OD600 value respectively.
According to experimental result, choose Cd
2+Concentration be 150 μ M as stress conditions, carried out the mensuration of growth curve.The thalline list spot that picking contains recombinant plasmid 37 ℃ of incubated overnight to the 10ml LB (containing Kan 50 μ g/ml).The 100ul bacterium is joined 10ml Cd
2+Concentration is in the LB substratum (containing Kan 50 μ g/ml, IPTG1mM) of 150 μ M, and 37 ℃, 200rpm shake cultivation, and is every at a distance from 2 hours mensuration OD600 values.
Experimental result shows that the BL21 (DE3) that expresses the DbsCDF gene is at 0-200 μ M Cd
2+Can normal growth under the concentration, and empty carrier is to impinging upon Cd
2+When surpassing 150 μ M, concentration just can not grow (seeing accompanying drawing 2).At 150 μ MCd
2+Under the concentration, the BL21 (DE3) that expresses the DbsCDF gene can grow in culturing process, and the growth of empty carrier contrast is suppressed (seeing accompanying drawing 3) always.The experimental result explanation, DbsCDF plays an important role to the defence of heavy metal cadmium.
SEQ?ID?NO：1
MAHDSHDHAGHTAGHHHTHAHSHAHGPANYGRAFAIGVALNLSFVLAEVVFGMIGHSLALLADAGHNMSDILGLLMAWGATALSRRQPSARFTYGLRSSSILVALANAVFLLVVTGGIAWEAIQRLLHPTPVASDIVIGVAAFGVAINTGTALLFMAGRKGDLNIRAAFLHMAGDAAVSLGVVLAGIGMLFTGWFWLDPTVSLLISAVIVIATWGLLRDSLNLTLHGVPEGIETQAVRAYLAALPDVAAVHDLHIWAMSTTETALTVHLVMPNGYPGDALLNRISNELQRRFRIDHPTLQVEAGDPAHPCALAPDHRV
SEQ?ID?NO：2
ATGGCACACGATTCTCATGACCATGCCGGCCATACCGCTGGTCACCATCATACCCACGCGCATAGCCATGCCCACGGTCCGGCAAACTACGGCCGCGCTTTTGCCATCGGGGTGGCGCTCAACCTGAGCTTCGTGCTGGCTGAGGTCGTGTTTGGGATGATTGGACATTCCCTGGCTTTGCTGGCGGATGCCGGACATAACATGAGTGACATCCTGGGCTTACTCATGGCTTGGGGCGCCACGGCACTGTCTCGCCGCCAGCCCTCGGCGCGCTTTACCTACGGGCTGCGCAGCTCGTCTATTCTGGTGGCTCTGGCCAATGCCGTATTCCTTTTGGTGGTGACTGGCGGGATCGCCTGGGAAGCGATTCAGCGCCTGCTGCATCCCACGCCCGTGGCAAGCGACATAGTCATTGGCGTCGCAGCCTTCGGAGTGGCGATCAATACTGGGACCGCCCTGCTGTTCATGGCGGGGCGCAAGGGCGATCTGAATATCCGGGCGGCATTTCTGCACATGGCGGGCGATGCCGCTGTATCGCTCGGGGTGGTCCTGGCGGGGATCGGGATGCTGTTTACTGGCTGGTTTTGGCTGGATCCCACGGTCAGCCTGCTCATCAGCGCGGTGATCGTGATCGCCACCTGGGGCTTGCTGAGGGACTCCCTCAACCTGACCCTGCACGGCGTGCCCGAAGGAATCGAAACCCAGGCGGTACGCGCCTATCTCGCCGCCCTGCCGGATGTGGCGGCGGTACATGACCTGCACATTTGGGCCATGAGCACCACCGAAACGGCGCTGACCGTGCATCTGGTCATGCCCAACGGCTACCCCGGAGACGCGCTCCTGAACCGGATCAGCAATGAATTGCAGAGGCGCTTTCGCATCGACCACCCCACCTTGCAGGTGGAGGCTGGCGATCCGGCCCACCCGTGCGCACTGGCGCCGGATCATCGGGTTTGA。
SEQUENCE?LISTING
< 110>Zhongshan University
< 120>a kind of supporing yang ion diffusion protein D bsCDF and encoding sox and application
<130>
<160> 4
<170> PatentIn?version?3.2
<210> 1
<211> 318
<212> PRT
< 213>artificial sequence
<400> 1
Met?Ala?His?Asp?Ser?His?Asp?His?Ala?Gly?His?Thr?Ala?Gly?His?His
1 5 10 15
His?Thr?His?Ala?His?Ser?His?Ala?His?Gly?Pro?Ala?Asn?Tyr?Gly?Arg
20 25 30
Ala?Phe?Ala?Ile?Gly?Val?Ala?Leu?Asn?Leu?Ser?Phe?Val?Leu?Ala?Glu
35 40 45
Val?Val?Phe?Gly?Met?Ile?Gly?His?Ser?Leu?Ala?Leu?Leu?Ala?Asp?Ala
50 55 60
Gly?His?Asn?Met?Ser?Asp?Ile?Leu?Gly?Leu?Leu?Met?Ala?Trp?Gly?Ala
65 70 75 80
Thr?Ala?Leu?Ser?Arg?Arg?Gln?Pro?Ser?Ala?Arg?Phe?Thr?Tyr?Gly?Leu
85 90 95
Arg?Ser?Ser?Ser?Ile?Leu?Val?Ala?Leu?Ala?Asn?Ala?Val?Phe?Leu?Leu
100 105 110
Val?Val?Thr?Gly?Gly?Ile?Ala?Trp?Glu?Ala?Ile?Gln?Arg?Leu?Leu?His
115 120 125
Pro?Thr?Pro?Val?Ala?Ser?Asp?Ile?Val?Ile?Gly?Val?Ala?Ala?Phe?Gly
130 135 140
Val?Ala?Ile?Asn?Thr?Gly?Thr?Ala?Leu?Leu?Phe?Met?Ala?Gly?Arg?Lys
145 150 155 160
Gly?Asp?Leu?Asn?Ile?Arg?Ala?Ala?Phe?Leu?His?Met?Ala?Gly?Asp?Ala
165 170 175
Ala?Val?Ser?Leu?Gly?Val?Val?Leu?Ala?Gly?Ile?Gly?Met?Leu?Phe?Thr
180 185 190
Gly?Trp?Phe?Trp?Leu?Asp?Pro?Thr?Val?Ser?Leu?Leu?Ile?Ser?Ala?Val
195 200 205
Ile?Val?Ile?Ala?Thr?Trp?Gly?Leu?Leu?Arg?Asp?Ser?Leu?Asn?Leu?Thr
210 215 220
Leu?His?Gly?Val?Pro?Glu?Gly?Ile?Glu?Thr?Gln?Ala?Val?Arg?Ala?Tyr
225 230 235 240
Leu?Ala?Ala?Leu?Pro?Asp?Val?Ala?Ala?Val?His?Asp?Leu?His?Ile?Trp
245 250 255
Ala?Met?Ser?Thr?Thr?Glu?Thr?Ala?Leu?Thr?Val?His?Leu?Val?Met?Pro
260 265 270
Asn?Gly?Tyr?Pro?Gly?Asp?Ala?Leu?Leu?Asn?Arg?Ile?Ser?Asn?Glu?Leu
275 280 285
Gln?Arg?Arg?Phe?Arg?Ile?Asp?His?Pro?Thr?Leu?Gln?Val?Glu?Ala?Gly
290 295 300
Asp?Pro?Ala?His?Pro?Cys?Ala?Leu?Ala?Pro?Asp?His?Arg?Val
305 310 315
<210> 2
<211> 957
<212> DNA
< 213>artificial sequence
<400> 2
atggcacacg?attctcatga?ccatgccggc?cataccgctg?gtcaccatca?tacccacgcg 60
catagccatg?cccacggtcc?ggcaaactac?ggccgcgctt?ttgccatcgg?ggtggcgctc 120
aacctgagct?tcgtgctggc?tgaggtcgtg?tttgggatga?ttggacattc?cctggctttg 180
ctggcggatg?ccggacataa?catgagtgac?atcctgggct?tactcatggc?ttggggcgcc 240
acggcactgt?ctcgccgcca?gccctcggcg?cgctttacct?acgggctgcg?cagctcgtct 300
attctggtgg?ctctggccaa?tgccgtattc?cttttggtgg?tgactggcgg?gatcgcctgg 360
gaagcgattc?agcgcctgct?gcatcccacg?cccgtggcaa?gcgacatagt?cattggcgtc 420
gcagccttcg?gagtggcgat?caatactggg?accgccctgc?tgttcatggc?ggggcgcaag 480
ggcgatctga?atatccgggc?ggcatttctg?cacatggcgg?gcgatgccgc?tgtatcgctc 540
ggggtggtcc?tggcggggat?cgggatgctg?tttactggct?ggttttggct?ggatcccacg 600
gtcagcctgc?tcatcagcgc?ggtgatcgtg?atcgccacct?ggggcttgct?gagggactcc 660
ctcaacctga?ccctgcacgg?cgtgcccgaa?ggaatcgaaa?cccaggcggt?acgcgcctat 720
ctcgccgccc?tgccggatgt?ggcggcggta?catgacctgc?acatttgggc?catgagcacc 780
accgaaacgg?cgctgaccgt?gcatctggtc?atgcccaacg?gctaccccgg?agacgcgctc 840
ctgaaccgga?tcagcaatga?attgcagagg?cgctttcgca?tcgaccaccc?caccttgcag 900
gtggaggctg?gcgatccggc?ccacccgtgc?gcactggcgc?cggatcatcg?ggtttga 957
<210> 3
<211> 31
<212> DNA
< 213>artificial sequence
<400> 3
cgcggatcca?tggcacacga?ttctcatgac?c 31
<210> 4
<211> 28
<212> DNA
< 213>artificial sequence
<400> 4
ccgctcgagt?caaacccgat?gatccggc 28
Claims (10)
1. a supporing yang ion diffusion albumen is characterized in that aminoacid sequence is shown in SEQ ID NO:1.
2. the described albumen of claim 1 is as the application of cadmium metal resistance-associated protein.
3. the application of the described albumen of claim 1 in administering Environmental Cadmium Pollution.
4. the nucleotide sequence of the described supporing yang ion diffusion protein molecular of coding claim 1 is characterized in that nucleotide sequence is shown in SEQ ID NO:2.
5. nucleotides sequence as claimed in claim 4 is listed in the application of administering in the Environmental Cadmium Pollution.
6. the expression vector that contains the said nucleotide sequence of claim 4.
7. a genetic engineering bacterium is characterized in that being obtained by the described expression vector transfection Escherichia coli of claim 6.
8. like the application of the said genetic engineering bacterium of claim 7 in administering Environmental Cadmium Pollution.
9. application as claimed in claim 8 is characterized in that the concentration of cadmium ions in the Environmental Cadmium Pollution is below the 200 μ M.
10. claim 1 or 3 described albumen or nucleotides sequence are listed in the application on the development of metallic cadmium resistant transgenic engineering product.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210079833 CN102617711B (en) | 2012-04-28 | 2012-04-28 | Cation diffusion assisting protein DbsCDF and encoding gene and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210079833 CN102617711B (en) | 2012-04-28 | 2012-04-28 | Cation diffusion assisting protein DbsCDF and encoding gene and application thereof |
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| Publication Number | Publication Date |
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| CN102617711A true CN102617711A (en) | 2012-08-01 |
| CN102617711B CN102617711B (en) | 2013-09-18 |
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| CN 201210079833 Expired - Fee Related CN102617711B (en) | 2012-04-28 | 2012-04-28 | Cation diffusion assisting protein DbsCDF and encoding gene and application thereof |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101555276A (en) * | 2009-05-25 | 2009-10-14 | 合肥工业大学 | Cadmium-resistant vegetable protein, coding gene and application thereof |
| CN101921779A (en) * | 2010-06-12 | 2010-12-22 | 上海大学 | Gene DvCRP2 with Cd2+ resistant and Cu2+ resistant functions, encoding protein and application thereof |
| CN102020706A (en) * | 2009-09-23 | 2011-04-20 | 中国科学院植物研究所 | Associated protein of biology cadmium resistance and coding gene of same |
| CN102417899A (en) * | 2011-12-19 | 2012-04-18 | 中山大学 | Metallic cadmium resistance associated protein KdpD and coding gene and application thereof |
-
2012
- 2012-04-28 CN CN 201210079833 patent/CN102617711B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101555276A (en) * | 2009-05-25 | 2009-10-14 | 合肥工业大学 | Cadmium-resistant vegetable protein, coding gene and application thereof |
| CN102020706A (en) * | 2009-09-23 | 2011-04-20 | 中国科学院植物研究所 | Associated protein of biology cadmium resistance and coding gene of same |
| CN101921779A (en) * | 2010-06-12 | 2010-12-22 | 上海大学 | Gene DvCRP2 with Cd2+ resistant and Cu2+ resistant functions, encoding protein and application thereof |
| CN102417899A (en) * | 2011-12-19 | 2012-04-18 | 中山大学 | Metallic cadmium resistance associated protein KdpD and coding gene and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| DELHAIZE E.等: "Genes Encoding Proteins of the Cation Diffusion Facilitator Family That Confer Manganese Tolerance", 《PLANT CELL》, vol. 15, no. 5, 31 May 2003 (2003-05-31), pages 1131 - 1142 * |
| GUFFANTI A.等: "An antiport mechanism fir a member of the cation diffusion facilitator family: divalent cations efflux in exchange for K+ and H+", 《MOLECULAR MICROBIOLOGY》, vol. 45, no. 1, 31 July 2002 (2002-07-31), pages 145 - 153 * |
| MUNKELT D.等: "The Chromosomally Encoded Cation Diffusion Facilitator Proteins DmeF and FieF from Wautersia metallidurans CH34 Are Transportors of Broad Metal Specificity", 《JOURNAL OF BACTERIOLOGY》, vol. 186, no. 23, 31 December 2004 (2004-12-31), pages 8036 - 8043 * |
| YOU X.Y.等: "Cation Diffusion Facilitator Family Transporter", 《UNIPROTKB/TREMBL》, 9 October 2011 (2011-10-09), pages 9 - 9 * |
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|---|---|
| CN102617711B (en) | 2013-09-18 |
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