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Various Expression System

Plant Expression System

Baculovirus Expression SystemYeast Expression System

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Plant Expression Systems High copy number plant

vector, pNosdcGUS, expressesthe GUS gene under thecontrol of the nopalinesynthase (pNos) promoter.

This vector may be used tomonitor virus production andtransfection efficiency.

Application:The GUS gene can be excisedusing the 5 XmaI and 3 SacI

sites to allow the insertion ofother genes to be expressedunder the same regulatoryelements in plants.

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The linked nucleic acid sequences further comprise multiplecloning sites. The nucleic acid segment encodes a plant virus movementprotein. The nucleic acid segment encodes a plant virus coat protein. Nucleic acid sequences further comprise a marker gene or aselectable marker gene. Nucleic acid sequences further comprise nucleic acid whichencodes a plant virus coat protein. Rather it can be said as nucleic acid sequences furthercomprise nucleic acid encoding a plant virus movementprotein. The linked nucleic acid sequences encode a fusionpolypeptide comprising the marker or selectable marker and

the plant virus coat protein or the plant virus movementrotein.

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A nucleic acid composition, comprising: (a) isolated RNA-1 offlock house virus; and (b) a recombinant RNA molecule

comprising linked RNA sequences comprising : RNA-1 or RNA-2 of flock house virus and RNA encoding a plant virus coatprotein. A method of expressing a gene product encoded by arecombinant RNA in a plant host cell, comprising:a. contacting the plant host cell with an amount of the

composition of isolated RNA-1 of flock house virus, arecombinant RNA comprising the vector.

b. detecting or determining whether the gene productencoded by the recombinant RNA is expressed.

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aculovirus Expression System

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Vectors pUCDM and pFBDM contain expressioncassettes comprising a central multiplication module(M), promoters (polh, p10), multiple cloning sites(MCS1, MCS2), and terminators (SV40, HSVtk) flankedby unique Pme I and Avr II (boxed) endonuclease sites.

pUCDM also contains the inverted repeat for Cre-loxP site-specific recombination ( LoxP ) and a

resistance marker for chloramphenicol. pFBDMcontains transposon elements ( Tn7L , Tn7R ) andresistance markers for ampicillin and gentamycin.

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Genes of interest are cloned into MCS1 or MCS2. ( b )Derivatives of pUCDM or pFBDM containingmultigene expression cassettes are assembled byexcising the expression cassette containing two genes(Gene A, Gene B) using Pme I and Avr II digestion.

Inserting the fragment generated into anothercassette containing more genes (Gene C, Gene D) viathe Bst Z17I/ Spe I or the Nru I/Spe I sites (bothpairs Pme I/ Avr II compatible) present in themultiplication module (M).

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The process is iterative. ( c) MultiBac is adaptable tocombinatorial applications of protein production.Example (1) shows multiple genes encoding forprotein subunits cloned in pUCDM using themultiplication module and inserted into MultiBacbacmid by Cre-loxP site-specific recombination.

In the same reaction, genes cloned singly in pFBDMencoding for a series of truncation variants of an

additional protein are inserted into the bacmid viaTn7 transposition. Example (2) shows differentenzymes cloned singly in pUCDM for insertionat LoxP with the purpose of modifying the proteinsalready expressed from Tn7 .

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Yeast Expression System

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