Selected article for: "expression vector and Klenow enzyme"
Title: Primary sequence domains required for the retention of rotavirus VP7 in the endoplasmic reticulum
  • Document date: 1988_11_1
    • ID: 63mxzwti_5
    Snippet: The deletion of the first initiation codon, and thus its accompanying hydrophobic domain, in the constructs A42-61/dhl, A43-61/dhl and A47-61/ dhl, was achieved by cutting plasmids A42-61, A43-61 and A47-61 (31) with Cla 1 (New England Biolabs, Beverly, MA) within the coding sequence of hl (Fig. 1) . Klenow enzyme (BRL, Gaitbersburg, MD) was then used to blunt the fragment ends and phosphorylated (22) Xho I linkers (New England Biolabs) were adde.....
    Document: The deletion of the first initiation codon, and thus its accompanying hydrophobic domain, in the constructs A42-61/dhl, A43-61/dhl and A47-61/ dhl, was achieved by cutting plasmids A42-61, A43-61 and A47-61 (31) with Cla 1 (New England Biolabs, Beverly, MA) within the coding sequence of hl (Fig. 1) . Klenow enzyme (BRL, Gaitbersburg, MD) was then used to blunt the fragment ends and phosphorylated (22) Xho I linkers (New England Biolabs) were added to the fragments generated. After Xho I digestion, the 1,000-bp VP7 coding fragment was isolated for each plasmid from Sea Plaque low melting temperature agarose (FMC Bioproducts, Rockland, ME) and cloned into the Xho I site of the expression vector pJCll9 (36), after calf alkaline phosphatase treatment (Boehringer Mannheim Biochemicals, Indianapolis, IN) of the vector.

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