TABLE 1.

Cloning strategy^a

Expression vector	Template for PCR	Primers		Cloning vector	Cloning site restriction fragment	Cloning site vector
		Forward	Reverse
pLipA.I	LipA cDNA	LipA5E/N	Lip3B/H	pAN52-10Notc	NcoIe-HindIII	NcoI-HindIII
pLipA.II	LipA cDNA	LipA5E/N	Lip3B/H	pAN52-6Notc	BssHII-HindIII	BssHII-HindIII
pGLA::LipAb	LipA cDNA	LipA5E/N	Lip3B/H	pAN56-2d	BssHII-BamHI	NarI-BglIIe
pMn1.I	Mnp1 cDNA	MNP15E/N	MNP13B/H	pAN52-10Not	NcoI-HindIII	NcoI-HindIII
pGLA::Mnp1b	Mnp1 cDNA	MNP1E/B	MNP13B/H	pAN56-2	BbsI-BamHI	NarI-BglIIe

^aFor each expression vector are indicated primers used for amplification of the LipA or Mnp1 cDNA clone and addition of cloning sites, recipient Aspergillus expression vector, and restriction sites used in the final cloning procedure. 

^bFor pGLA::LipA and pGLA::Mnp1, the Kex2 site containing linker NarI-NVISKR-BssHII was used to join vector and insert molecules. 

^cThese vectors are derivatives from pAN52-4 (Punt, unpublished) with improved cloning sites. 

^dGenBank accession number z32690. 

^ePartial digestion.