Restriction digests of the clone give the following sizes (kb): EcoRI/HindIII--2.7, 0.7; EcoRI--3.4; HindIII--3.4; PstI--3.4; BamHI--3.4. The ApaI fragment extending from nt 58 in the pIBI25 polylinker to nt 569 in the 3' noncoding region has been used as a probe. This contains the complete 441 bp coding sequence as well as portions of the 5' (44 bp) and 3' (15 bp) noncoding regions. The coding sequence extends from nt 113 to nt 554 of the insert. Nucleotides 1-69 are derived from the pIBI25 polylinker. The AvaII polymorphism results from a silent transversion at the third position of codon +86. Derived from a plasmid constructed by cloning a genomic AccI fragment into the HincII site of pIBI25. The EcoRV insert thus contains some sequences at the 5' end derived from the pIBI25 polylinker. |
