Invitrogen™

Multi-Copy Pichia Expression Kit

The Multi-Copy Pichia Expression Kit contains the pPIC3.5K, pPIC9K, and pAO815 vectors for production and selection of Pichia strains that자세히 알아보기

The Multi-Copy Pichia Expression Kit contains the pPIC3.5K, pPIC9K, and pAO815 vectors for production and selection of Pichia strains that contain more than one copy of the gene of interest. In many cases, increased copy number has led to increased expression levels.

*pPIC9K and pPIC3.5K
*The pPIC9K and pPIC3.5K vectors carry the kanamycin resistance gene that confers resistance to Geneticin™ Reagent in Pichi_a. Spontaneous generation of multiple insertion events can be identified by resistance to increased levels of Geneticin™ Reagent. _Pichia transformants are selected on histidine-deficient medium and screened for their level of resistance to Geneticin™ Reagent. The ability to grow in high concentrations of Geneticin™ indicates that multiple copies of the kanamycin resistance gene and the gene of interest are integrated into the genome (1). The pPIC9K vector directs secretion of expressed proteins while proteins expressed from pPIC3.5K remain intracellular.

*pAO815
*pAO815 is a Pichia expression vector designed to clone multiple copies of a gene into a single vector. The vector contains a Bgl II site upstream of the 5´ AOX1 gene and a unique Ba_mH I site downstream of the 3´ _AOX1 transcription termination (TT) signal. Four steps are required to generate multiple copies of the gene of interest:
1. The gene is cloned into the unique Ec_oR I site in the vector.
2. The construct is digested with _Ba_mH I and _Bgl II to release the “expression cassette” containing the AOX1 promoter, gene of interest, and 3´ AOX1 TT.
3. Concatemers of the expression cassette are generated by ligation _in vitr_o.
4. The multiple copies are inserted back into the pAO815 vector and transformed into _Pichi_a.

These vectors are also available separately.

사양

항생제 내성 박테리아Ampicillin (AmpR), Gentamicin (GmR)

박테리아 또는 효모 균주KM71

클로닝 방법Restriction Enzyme/MCS

셀 유형Yeast Cells

형식Kit

프로모터AOX1

종P. pastoris

VectorpPIC

발현 메커니즘Cell-Based Expression

발현 시스템Yeast

용도(애플리케이션)Protein Expression

제품 유형Expression Kit

단백질 태그Untagged

수량1 Kit

선택 제제(진핵)Ampicillin (AmpR), Gentamicin (GmR)

Unit SizeEach