Title: Membrane protein retention in the yeast Golgi apparatus: dipeptidyl aminopeptidase A is retained by a cytoplasmic signal containing aromatic residues Document date: 1993_6_2
ID: 0pz80zbg_15
Snippet: Deletions within the cytoplasmic domain of DPAP A (see Fig. l ) were constructed via oligonucleotide mutagenesis of pCJR71. To incorporate these deletions into the A-ALP fusion protein, SacI-Bglll fragments carrying the appropriate deletions were fused to the BglII-EcoRI fragment from pSN14 and both fragments were inserted into the SacI-EcoRI sites of pSEYC68 (a URA3-based, CEN plasmid; Emr et al., 1983 ) via a three-way ligation. The following i.....
Document: Deletions within the cytoplasmic domain of DPAP A (see Fig. l ) were constructed via oligonucleotide mutagenesis of pCJR71. To incorporate these deletions into the A-ALP fusion protein, SacI-Bglll fragments carrying the appropriate deletions were fused to the BglII-EcoRI fragment from pSN14 and both fragments were inserted into the SacI-EcoRI sites of pSEYC68 (a URA3-based, CEN plasmid; Emr et al., 1983 ) via a three-way ligation. The following is a list of the resulting plasmids and encoded protein products: pSN25 (A2-51-A-ALP), pSN32 (A2-80-A-ALP), pSN34 (A2-100-A-ALP), pSN35 (A109-116-A-ALP), pSN27 (A68-106-A-ALP), pSN60 (A85-106-A-ALP), pSN41 (Ag5-92-A-ALP), pSN42 (A92-99-A-ALP), and pSN43 (A99-106-A-ALP), where the deletion endpoints are indicated (e.g., A2-51 is missing residues 2 through 51).
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