pDL2产黄青霉基因整合质粒载体(Penicillium chrysogenum) BioVector NTCC质粒载体菌种细胞基因保藏中心

pDL2产黄青霉基因整合质粒载体(Penicillium chrysogenum) BioVector NTCC质粒载体菌种细胞基因保藏中心
Two plasmids, pDL2 and pDL10, were constructed for double
crossing over experiments (Fig. 3). Plasmid pDL2 contained
the same 2.1-kb DNA fragment internal to lys2 used in
pDL1, with a 1.3-kb EcoRV internal fragment of the lys2 gene
replaced by a 1.5-kb XhoI-EcoRI fragment containing the
phleomycin resistance gene (ble) under the control of the Acremonium
chrysogenum pcbC promoter. A linear 2.3-kb XhoIBamHI
fragment from pDL2, in which two regions (0.36 and
0.43 kb) homologous to the 59 and 39 regions of lys2 flanked the
ble gene, was used for the transformation.
In plasmid pDL10 the pyrG gene was inserted to inactivate
the lys2 gene, and in addition an internal PstI-EcoRV fragment
of 200 bp was removed to avoid the reversion of the lys2
mutation by further recombination processes. A linear 8.8-kb
NotI-KpnI fragment from pDL10, in which two regions (4.3
and 3 kb) homologous to the lys2 region are located at both
sides of the pyrG gene, was used for the transformation.
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