Restriction digests of the clone give the following sizes (kb): BamHI--7.8, 3.8; HindIII--6.8, 4.7. The clone appears to be unstable. We recommend that the ATCC-provided stock be streaked for single colonies and that a colony with the correct restriction pattern be selected. This culture was previously deposited as ATCC 53063. An Escherichia coli/Bacillus spp. shuttle expression vector for the construction and secretion of fusion proteins. Contains the alkaline phosphatase gene of Escherichia coli. pBs81/6 (Bacillus replication) was cut with HindIII and cloned into the HindIII site. A 1.1 kb EcoRI fragment with Bacillus licheniformis alpha-amylase regulatory and signal sequences was cloned into pUC9 cut with EcoRI/SmaI. A 3.0 kb BamHI/HindIII fragment (Escherichia coli phoA minus signal sequences and the first 5-13 aa) was inserted. |
