The 3' end PCR products were purified and ligated into pMD-18T cloning vector
(Takara Biotechnology, China).
3Kb) was gel purified and cloned into a cloning vector
(pUC29) at NdeI and Xhol restriction sites.
pUC18 and IGF-1 gene were purchased from GenScript (USA).
After moving to National Institutes of Health as Visiting Scientist in 1983, he developed a high-efficiency transfection method for delivering cDNA libraries into mammalian cells and cDNA cloning vectors
that enable selection on stable cDNA expression.
In this paper, we report PCR amplification and cloning of a cellulase gene into PTZ57R/T cloning vector
from genomic DNA of an indigenous strain of Bacillus sp.
After purification of PCR product from the gel, both fragment and STV28 cloning vector
were doubled digested with HindIII and PstI.
The amplified product was purified by PCR purification kit following manufacturer's instructions (Qiagen, Germany) and cloned first in the TA cloning vector
pTZ57R (Fermantas) followed by subcloning in the yeast shuttle expression vector pGal426 that contains the galactose inducible promoter Gall and uracil synthesizing gene (ura3) as selection marker (12,13).
asburiae MS7 followed by ligation of DNA fragments into the BamHI-site of cloning vector
pSPPO1 contained a BamHI fragment and pAPPO1 a SacI fragment of cloning vector
between the promoter and the sense transgene, and between the antisense transgene and terminator respectively.
The expression cassette of the cloning vector
is illustrated in supplemental file 4 in the online Data Supplement.
A cDNA of the target transcript is cloned in an appropriate cloning vector
Each PCR fragment was inserted into the cloning vector
TSAP catalyzes the removal of 5' phosphate groups from DNA, thus preventing the recircularization and religation of linearized cloning vector
DNA during ligation.
The amplifications were ligated in pTZ57R/T cloning vector
PCR amplicon of CVMV-CP gene with 531 bp was cloned into pTZ57R/T (2886 kb) a cloning vector