scholarly journals VERIFIKASI cDNA T29 SEBAGAI KANDIDAT GEN PENGKODE PROTEIN Toxoplasma gondii DENGAN METODE SDS PAGE

2005 ◽  
Vol 11 (1) ◽  
pp. 61-66
Author(s):  
Ira Djajanegara ◽  
Wayan Artama ◽  
Retno Lestari ◽  
Sabar Pambudi
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Plasmid ◽  
Restriction Site ◽  
Standard Procedure ◽  
Pcr Amplification ◽  
Gene Encoding ◽  
E Coli ◽  
Gene Coding ◽  
Sds Page ◽  
Encoding Protein

The process of cDNA construction from mRNA isolated from Toxoplasma gondii has been done. There were 7 candidates cDNA which one of them is called T29. Since Toxoplasma gondii is the cause of toxoplasmosis infection, cloning the gene encoding protein from this parasite provides an important tool for developing diagnostic kit for detection of toxoplasmosis. Digestion of the cDNA T29 with EcoRI which is the restriction site where the cDNA was inserted yielded a 1.862 bp fragment. The fragment was subcloned into E. coli expression vector pMal-p2x and transformed into E.coli strain TB1. Colonies of TB1 were grown on ampicillin plates and the recombinant plasmid was extracted using the standard procedure. The plasmid was digested using EcoRI and PstI, checked by PCR amplification using malE and M13/pUC primers. The recombinant plasmid was expressed in TB1 and the protein extracted was ran in SDS PAGE to observe the presence of the expressed protein. Based on the data from this experiment, there was no expression result of the expressed cDNA which was confirm by the PCR result. Therefore, it was concluded that cDNA T29 was not carrying the gene coding for protein from parasite Toxoplasma gondii.

scholarly journals Cloning and Sequencing cDNA Encoding for Rhoptry-2 Toxoplasma Gondii Tachyzoite Local Isolate

2015 ◽  
Vol 10 (2) ◽  
Author(s):  
Wayan T. Artama ◽  
Yulia Sari ◽  
Didik Tulus Subekti ◽  
Soenarwan Hery Poerwanto ◽  
Jarot Subandono
Keyword(s):  
Toxoplasma Gondii ◽  
Recombinant Plasmid ◽  
Messenger Rna ◽  
Secretory Protein ◽  
Gene Encoding ◽  
Isolation System ◽  
Complementary Dna ◽  
E Coli ◽  
Local Isolate ◽  
Cell Target

Rhoptry protein belongs to an excretory and secretory antigens (ESAs) that play an important role during activepenetration of parasite into the cell target. This protein an able Toxoplasma gondii to actively penetrate targetedcell, meanwhile ESAs protein stimulates intracellular vacuole modification. It is, therefore, after the parasitesuccessfully enter the cell target then Granule (GRA) proteins are responsible for the formation of parasitophorusvacuole, which is protect the fusion with other intracellular compartments such as lysosomal vacuole. Consequently,this parasite is being able to survive and multiply at the cell target. The current study was aimed to clone andsequens cDNA encoding for ROP-2 of local isolated T. gondii tachizoite through DNA recombinant technique.Total ribonucleic acid (RNA) was isolated from tachyzoites of local isolated T. gondii that were grown up in Balb/c mice. Messenger RNA was isolated from total RNA using PolyAtract mRNA Isolation System. Messenger RNA wasused as a template for synthesis cDNA using Riboclone cDNA Synthesis System AMV-RT. EcoRI adaptor fromRiboclone EcoRI Adaptor Ligation System was added to Complementary DNA and than ligated to pUC19. Recombinantplasmid was transformed into E. coli (XL1-Blue). The transformed E. coli XL-1 Blue were plated on LB agarcontaining X-Gal, IPTG and ampicillin. Recombinant clones (white colony) were picked up and grown up in theLB medium at 37oC overnight. Expression of recombinant protein was analysed by immunoblotting in order toidentify cDNA recombinant wich is express ESA of T. gondii local isolate. Recombinant plasmid were isolatedusing alkalilysis method and were elektroforated in 1% agarose gel. The isolated DNA recombinant plasmid wascut using Eco RI and then sequenced through Big Dye Terminator Mix AB1 377A Sequencer using M13 Forward andM13 Reverse primers. The conclusion of this results showed that the recombinant clone was coding for excretoryand secretory protein which has molecular weight of 54 kDa. The DNA alignments of sequence from the clonedgene showed 97% homology with gene encoding for ROP-2 of T. gondii RH isolate.Keywords: Toxoplasma gondii, tachizoite, ESA, complementary DNA, ROP2

scholarly journals Expression of Recombinant Fusion Protein of Newcastle Disease Virus from Escherichia coli Plasmid Clone C-2a by In-vitro Cell-free Protein Expression System

2020 ◽  
Vol 20 ◽  
pp. 04004
Author(s):  
Ahmad Pandu Satria Wiratama ◽  
Aris Haryanto
Keyword(s):  
Protein Expression ◽  
Newcastle Disease ◽  
Recombinant Plasmid ◽  
Expression System ◽  
Free Protein ◽  
F Protein ◽  
E Coli ◽  
Protein Expression System ◽  
Sds Page

Newcastle Disease Virus (NDV) is an infectious disease that infect many kinds of wild and domesticated birds. Infection of NDV become a massive problem for poultry industry around the world especially in Indonesia. Vaccination is an effort to prevent the infection of NDV in poultry. NDV vaccine that used in Indonesia is a conventional life vaccine from LaSota and B1 strains. These type of vaccine is 21%-23% genetically distinct with the virus that spread in the environment. The antibody protection provided by the vaccine is not effective. Therefore, vaccination with new local NDV strain is needed to prevent the NDV infection in Indonesia. The previously study research reported that the local isolate of NDV from Kulon Progo, Indonesia has been isolated. Fusion (F) protein encoding gene that has been inserted into pBT7-N-His expression p lasmid which isolated from clone C-2a of E. coli, then it was expressed by the Cell-free protein expression system. The aim of this study was to confirm whether clone C-2a of E.coli carrying a recombinant plasmid pBT7-N-His-Fusion NDV and to express a recombinant F protein of NDV in-vitro from expression plasmid by cell-free protein expression system. This work started by detection of recombinant plasmid pBT7-N-His-Fusion NDV by DNA plasmid extraction followed by agarose gel electrophoresis. The recombinant F protein was in-vitro expressed by cell-free protein expression kit. The expressed F protein of NDV then was visualized by SDS-PAGE and Westernblott to analyse the expression of NDV recombinant F protein. It confirmed that clone C-2a of E. coli contained plasmid pBT7-N-His (4.001 bp) inserted by recombinant F protein of NDV gene (642 bp). The visualisation of expressed recombinant F protein by SDS-PAGE and Westernblott showed the NDV recombinant F protein was a specific protein fragment with molecular weight of 25,6 kDa..

scholarly journals S-Adenosylmethionine Transport in Rickettsia prowazekii

2003 ◽  
Vol 185 (10) ◽  
pp. 3031-3035 ◽  
Author(s):  
Aimee M. Tucker ◽  
Herbert H. Winkler ◽  
Lonnie O. Driskell ◽  
David O. Wood
Keyword(s):  
Evolutionary Relationship ◽  
Host Cells ◽  
Transport Systems ◽  
Rickettsia Prowazekii ◽  
Gene Encoding ◽  
E Coli ◽  
Obligate Intracellular ◽  
Gene Coding ◽  
Epidemic Typhus ◽  
Methionine Transport

ABSTRACT Rickettsia prowazekii, the causative agent of epidemic typhus, is an obligate, intracellular, parasitic bacterium that grows within the cytoplasm of eucaryotic host cells. Rickettsiae exploit this intracellular environment by using transport systems for the compounds available in the host cell's cytoplasm. Analysis of the R. prowazekii Madrid E genome sequence revealed the presence of a mutation in the rickettsial metK gene, the gene encoding the enzyme responsible for the synthesis of S-adenosylmethionine (AdoMet). Since AdoMet is required for rickettsial processes, the apparent inability of this strain to synthesize AdoMet suggested the presence of a rickettsial AdoMet transporter. We have confirmed the presence of an AdoMet transporter in the rickettsiae which, to our knowledge, is the first bacterial AdoMet transporter identified. The influx of AdoMet into rickettsiae was a saturable process with a KT of 2.3 μM. Transport was inhibited by S-adenosylethionine and S-adenosylhomocysteine but not by sinfungin or methionine. Transport was also inhibited by 2,4-dinitrophenol, suggesting an energy-linked transport mechanism, and by N-ethylmaleimide. AdoMet transporters with similar properties were also identified in the Breinl strain of R. prowazekii and in Rickettsia typhi. By screening Escherichia coli clone banks for AdoMet transport, the R. prowazekii gene coding for a transporter, RP076 (sam), was identified. AdoMet transport in E. coli containing the R. prowazekii sam gene exhibited kinetics similar to that seen in rickettsiae. The existence of a rickettsial transporter for AdoMet raises intriguing questions concerning the evolutionary relationship between the synthesis and transport of this essential metabolite.

scholarly journals Cloning, Sequencing, and Expression of the Gene Encoding Cyclic 2,3-Diphosphoglycerate Synthetase, the Key Enzyme of Cyclic 2,3-Diphosphoglycerate Metabolism in Methanothermus fervidus

1998 ◽  
Vol 180 (22) ◽  
pp. 5997-6004 ◽  
Author(s):  
Karl Matussek ◽  
Patrick Moritz ◽  
Nina Brunner ◽  
Christoph Eckerskorn ◽  
Reinhard Hensel
Keyword(s):  
Substrate Affinity ◽  
Kinetic Properties ◽  
Gene Encoding ◽  
Enzyme Preparations ◽  
E Coli ◽  
Gene Coding ◽  
Key Enzyme ◽  
Methanothermus Fervidus ◽  
Synthetic Reaction ◽  
Sequencing And Expression

ABSTRACT Cyclic 2,3-diphosphoglycerate synthetase (cDPGS) catalyzes the synthesis of cyclic 2,3-diphosphoglycerate (cDPG) by formation of an intramolecular phosphoanhydride bond in 2,3-diphosphoglycerate. cDPG is known to be accumulated to high intracellular concentrations (>300 mM) as a putative thermoadapter in some hyperthermophilic methanogens. For the first time, we have purified active cDPGS from a methanogen, the hyperthermophilic archaeon Methanothermus fervidus, sequenced the coding gene, and expressed it in Escherichia coli. cDPGS purification resulted in enzyme preparations containing two isoforms differing in their electrophoretic mobility under denaturing conditions. Since both polypeptides showed the same N-terminal amino acid sequence and Southern analyses indicate the presence of only one gene coding for cDPGS in M. fervidus, the two polypeptides originate from the same gene but differ by a not yet identified modification. The native cDPGS represents a dimer with an apparent molecular mass of 112 kDa and catalyzes the reversible formation of the intramolecular phosphoanhydride bond at the expense of ATP. The enzyme shows a clear preference for the synthetic reaction: the substrate affinity and the V max of the synthetic reaction are a factor of 8 to 10 higher than the corresponding values for the reverse reaction. Comparison with the kinetic properties of the electrophoretically homogeneous, apparently unmodified recombinant enzyme from E. coli revealed a twofold-higher V max of the enzyme from M. fervidus in the synthesizing direction.

scholarly journals EKSPRESI GEN PENYANDI b-XILOSIDASE DALAM SISTEM pHIS1525/ Bacillus megaterium MS941

2008 ◽  
Vol 13 (2) ◽  
pp. 157-161
Author(s):  
Sri Sumarsih ◽  
Ni Nyoman Tri Puspaningsih ◽  
Sofijan Hadi ◽  
Ami Soewandi J.S.
Keyword(s):  
Bacillus Megaterium ◽  
Recombinant Plasmid ◽  
Culture Medium ◽  
Protein Band ◽  
Extracellular Protein ◽  
Protoplast Transformation ◽  
E Coli ◽  
Sds Page ◽  
Xylosidase Activity

The aim of this research was to express the β-xylosidase gene in the pHIS1525 or Bacillus megaterium MS941 system. The xyl gene was amplified from pTP510 and cloned into pHIS1525 in E. coli DH10b. The recombinant plasmid was transformed into B. megaterium MS941 by protoplast transformation. Transformants were selected by growing the recombinant B. megaterium MS941 on solid LB medium containing tetracycline (10 μg/ml). The expression of β-xylosidase was assayed using 0.2 percent methylumbelliferyl-β-D-xyloside (MUX) and the proteins were analyzed by SDS-PAGE method. The b-xilosidase activity was determined toward p-nitrophenyl-b-Dxylopyranoside (pNPX) as a substrate and p-nitrofenol releasing was measured by UV/Vis spectrophotometer at λ 405 nm. This research showed that recombinant B. megaterium MS941 expressed the β-xylosidase gene (xyl) and secreted it into the culture medium. The SDS-PAGE analysis of extracellular protein (culture medium) showed a 60,0 kD protein band. The recombinant Bacillus megaterium MS941 expressed and secreted the β-xilosidase into culture medium 5 hours after adding 5 percent xylose. The b-xylosidase activity was 0.441 unit/ml toward pNPX as a substrate.

scholarly journals Clonning and expression of recombinant carbapenemase KPC-2 enzyme in E. coli cytoplasm

2016 ◽  
Vol 19 (2) ◽  
pp. 27-37
Author(s):  
Phuong Nhat Tran ◽  
Phuong Thi Kim Huynh ◽  
Trang Thi Phuong Tran ◽  
Thuoc Linh Tran ◽  
Van Hung Pham
Keyword(s):  
Klebsiella Pneumoniae ◽  
Clinical Research ◽  
Affinity Chromatography ◽  
Recombinant Plasmid ◽  
Broth Culture ◽  
E Coli ◽  
Recombinant Vector ◽  
Carbapenem Resistant ◽  
Sds Page ◽  
Encoding Genes

Production of KPC-type carbapenemase is the most common carbapenem resistant mechanism in Klebsiella pneumoniae. The expression level of KPC in these strains is different and is mostly required other mechanisms to reach the higher resistant level such as porin lost or co-expression of extended spectrum β-lactamase (ESBL). To better understand the expression of KPC enzyme, the KPC-2 encoding genes from clinical isolated K. pneumoniae were cloned into pET28a plasmid. The recombinant plasmids containing of kpc-2 gene were subsequently transformed into E. coli OmniMax and were screened in kanamycine added LB media to select E. coli possessing of recombinant plasmid. Carbapenemase activity in the broth culture was checked in LB broth supplemented with 4 µg/mL of ertapenem and the expression induced with IPTG was checked by SDS-PAGE method. The results showed that this recombinant vector was capable of effective expression of KPC-2 protein in E. coli and this strain could be grown in LB broth supplemented with 4 µg/mL of ertapenem. A half of the target protein was soluble in the supernatant however it could be successfully collected from a HistrapHP affinity chromatography column. The result of this report is one of resources for further studies and applications of this KPC-2 protein in clinical research.

Operon Construction Containing alsS, ilvC, ilvD and kivd Genes in Esch-erichia coli for Isobutanol Production from Glucose

2020 ◽  
Vol 25 (1) ◽  
pp. 11-17
Author(s):  
Gabriel Kennardi ◽  
◽  
Maelita Moeis ◽  
Andreas Andreas ◽  
◽  
...  
Keyword(s):  
Biosynthetic Pathway ◽  
Acetolactate Synthase ◽  
Gas Chromatography Mass Spectrometry ◽  
Selected Ion Monitoring ◽  
E Coli ◽  
Lac Operator ◽  
Gene Coding ◽  
Sds Page ◽  
Weak Expression ◽  
Lac Promoter

Isobutanol is a biofuel considered to be a potential gasoline substitute. However, isobutanol production is difficult because there is no native organism that can produce isobutanol. A biosynthetic pathway to produce isobutanol had been designed to utilize pyruvate produced from glucose breakdown by glycolysis in Escherichia coli (E. coli). This biosynthetic pathway con-sists of acetolactate-synthase (ALS), ketol-acid reductoisomerase (KARI), dihydroxy-acid dehydratase (DHAD), alpha-ketoisovalerate decarboxylase (KDC) and alcohol dehydrogenase (ADH) enzymes. Since E. coli does not have ALS and KDC, the genes coding for the protein is needed to be cloned and overexpressed in E. coli. KARI and DHAD were overexpressed to increase the accumulation of keto acid to increase isobutanol production. Plasmid contains an operon controlled by lac pro-moter and lac operator consisting of alsS (coded ALS from Bacillus subtilis), ilvC (coded KARI from E. coli MG1655) and ilvD (coded DHAD from E. coli MG1655) genes, obtained from previous research, and operon sequences have been confirmed by DNA sequencing. kivd gene (coding KDC from Lactococcus lactis) was obtained from iGEM 2013 kit. kivd was amplified by PCR and inserted into pJET 1.2 blunt. kivd gene was then added into 3’ end of previous operon using restriction-ligation tech-nique. The plasmid constructed was then transfered into E. coli DH5α using heat shock. The recombinant genes were expressed using IPTG (isopropyl-β-D-1-thiogalactopyranoside) induction. The SDS PAGE results were inconclusive, however isobutanol was detected by Gas Chromatography Mass Spectrometry – Selected Ion Monitoring (GC-MS-SIM) from 48 hours fermenta-tion culture at 30 oC (1,17%). An operon regulated by the lac promoter-operator containing four genes for the biosynthesis of isobutanol has been constructed and cloned in E. coli. The isobutanol production was not optimal due to weak expression and repression by glucose, which was used as substrate.

scholarly journals CLONING AND EXPRESSING HG-CSF (HUMAN GRANULOCYTE COLONY STIMULATING FACTOR) BEING FUSED WITH FERRITIN HEAVY-CHAIN IN E. COLI

2009 ◽  
Vol 12 (9) ◽  
pp. 47-53
Author(s):  
Hieu Thi Phuong Nguyen ◽  
Linh Thuy Lien ◽  
Thuoc Linh Tran
Keyword(s):  
Heavy Chain ◽  
Cloning And Expression ◽  
Form Expression ◽  
Gene Encoding ◽  
E Coli ◽  
Ferritin Heavy Chain ◽  
Tev Protease ◽  
Sds Page ◽  
Human Ferritin ◽  
Stimulating Factor

G-CSF is a cytokine that stimulates the proliferation, differentiation, function of mature neutrophils and is generally used for treatment of neutropenia in cancer patients under chemotherapy. In this study, we report results on the cloning and expression of hG-CSF being fused with the heavy-chain (FTN-H) of human ferritin, which had been showed to be capable of facilitate the folding of several human protein expressed in E. coli. The hg-csf gene and gene encoding FTN-H were inserted into plasmid pET28a to form expression vector PET-FHG. 6xHis tag and TEV sequence (being recognized and cleaved by TEV protease) was added between of G-CSF and FTN-H to facilitate G-CSF purification and recovery afterwards. PET-FHG was transformed into E. coli BL21(DE3). The expression of hG-CSF was induced by IPTG and confirmed by SDS-PAGE and Western blot using anti-hG-CSF antibody.

scholarly journals SUB-CLONING OF GENES ENCODINGCYTHOCHROME P450 MONOOXYGENASE (CYP71AVI) INTOEXPRESSION VECTOR IN ESCHERICHIA COLI

2017 ◽  
Vol 10 (14) ◽  
pp. 69
Author(s):  
Imam Adi Wicaksono ◽  
Tresna Lestari ◽  
Evi Umayah Ulfa ◽  
Catur Riani ◽  
Elfahmi Elfahmi
Keyword(s):  
Escherichia Coli ◽  
Restriction Site ◽  
Restriction Analysis ◽  
Sequencing Analysis ◽  
Gene Encoding ◽  
P450 Monooxygenase ◽  
Recombinant Vector ◽  
Sds Page ◽  
Key Enzyme ◽  
Artemisinin Biosynthesis

Objective: Cytochrome P450 monooxygenase (CYP71AVI) is a key enzyme involved in the artemisinin biosynthesis pathway.In this research, sub-cloning gene encoding CYP71AVI into pETDUET1 vector in Escherichia coli has been done and then the expression products characterized with SDS-PAGE.Methods: Gene construction started with sub-cloning of cyp71avi gene from pJexpress401_cyp into pETDUET1 through restriction site NdeI and XhoI to get pETDUET1_cyp. Overproduction of CYP71AVI at temperature 37 °C has conducted by IPTG induction.Results: Confirmation of the recombinant vector pETDUET1_cyp was done by migration, restriction site and sequencing analysis. The result of pETDUET1_cyp restriction analysis with XhoI restriction enzyme showed one DNA band with experimental size 6585 bp.The CYP71AVI protein has been produced and characterized with SDS-PAGE method. Based on experimental calculation from SDS-PAGE analysis obtained molecular weight of CYP71AVI band was 57.55 kDa.Conclusion: Construction of gene encoding CYP71AVI into pETDUET1 as the co-expression vector in Escherichia colihas been succesfully and confirmed by migration, restriction site and sequencing analysis. The result of overproduction showed protein bands on SDS-PAGE analysis indicated as CYP71AVI. 

scholarly journals Ekspresi gen penyandi ACCase subunit biotin karboksilase dari mesokarp buah kelapa sawit pada Escherichia coli Expression of gene encoding ACCase subunit biotin carboxylase from oil palm fruit mesocarp in Escherichia coli

2016 ◽  
Vol 82 (1) ◽  
Author(s):  
Asmini BUDIANI
Keyword(s):  
Escherichia Coli ◽  
Oil Palm ◽  
Biotin Carboxylase ◽  
Acetyl Coa Carboxylase ◽  
Acetyl Coa ◽  
Gene Encoding ◽  
Palm Fruit ◽  
E Coli ◽  
Target Dna ◽  
Sds Page

Abstract Production of palm oil could be increased, one of which is by increasing oil content in the mesocarp of oil palm. This might be done by increasing the activity of key enzymes of the oil biosynthesis pathway in the oil palm mesocarp. Acetyl-CoA carboxylase has been reported as the enzyme that plays important role in oil accumulation in the oil palm mesocarp. Gene encoding one subunit of ACCase, biotin carboxylase (BC) had been isolated from oil palm mesocarp and cloned in E. coli. This reseach was aimed to examine the expression of the cloned BC gene in the E. coli. The cloned cDNA encoding BC was reisolated from recom-binant E. coli by PCR using spesific primers. The PCR products were verified in the agarose gel, and then ligated to pTrcHis-TOPO expression vector. The ligation product, recombinant vector pTrcHis-TOPO/BC, was introduced into E. coli XL1-Blue. Recombinant colonies grew in the selection media were analyzed using PCR to confirm the existent of the target DNA.  The colonies, which have been confirmed to contain target DNA were then subcultured in the LB media, for extraction of total protein. The protein extract was then analyzed quantitatively by Lowry method, and qualitatively by electrophoresis on SDS polyacrylamide gel. The result showed that recombinant plasmid pTrcHis-TOPO/BC has been successfully inserted into E. coli XL-1 Blue. SDS-PAGE analysis of the extracted protein showed that recombinant E. coli produced specific protein with MW of about 43 kDa, much higher compared with that of untransformed E. coli. This results demonstrate that  the cloned BC was strongly expressed in E. coli Abastrak Produksi minyak sawit dapat ditingkatkan, salah satu-nya dengan meningkatkan rendemen minyak. Hal ini dapat dilakukan dengan cara  meningkatkan aktivitas enzim kunci biosintesis minyak pada mesokarp buah sawit. Acetyl-CoA carboxylase telah dilaporkan merupakan enzim yang berperan penting dalam akumulasi minyak pada mesokarp kelapa sawit. Pada penelitian sebelumnya, gen penyandi salah satu subunit ACCase, yaitu biotin carboxylase (BC), telah diisolasi dari jaringan mesokarp kelapa sawit dan diklon pada E.coli. Tujuan penelitian ini adalah untuk menguji ekspresi gen tersebut pada E. coli. cDNA penyandi BC diisolasi kembali dari E. coli rekombinan dengan PCR menggunakan pasangan primer spesifik. Hasil isolasi diveri-fikasi pada gel agarose, kemudian diligasikan dengan vektor ekspresi  pTrcHis-TOPO.  Vektor  rekombinan (pTrcHis-TOPO/BC) hasil ligasi diintroduksikan ke dalam E. coli XL1-Blue. Koloni rekombinan yang tumbuh pada media seleksi dianalisis menggunakan PCR untuk mengkonfirmasi ada tidaknya sisipan DNA target. Koloni yang  terbukti mengandung sisipan DNA target dikulturkan pada media LB kemudian protein total diekstrak dari kultur E. coli dan dianalisis dengan elektroforesis SDS-PAGE. Hasil PCR koloni menunjukkan bahwa transformasi E. Coli XL-1 Blue menggunakan konstruk vektor rekombinan pTrcHis-TOPO/ BC berhasil baik. Analisis SDS-PAGE dari ekstrak protein menunjukkan bahwa E. coli rekombinan menghasilkan protein dengan berat molekul sekitar 43 kDa yang inten-sitasnya jauh lebih tinggi dibandingkan dengan protein yang sama yang dihasilkan oleh E. coli  yang tidak ditrans-formasi. Hal ini membuktikan bahwa gen penyandi BC dalam vektor pTrcHis-TOPO dapat diekspresikan dengan kuat pada   E. coli.

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