Isolation, characterization, and expression of cDNAs encoding murine alpha-mannosidase II, a Golgi enzyme that controls conversion of high mannose to complex N-glycans

Chromosome Mapping of the Human Man II Gene Comparison of the murine Man II cDNA with the sequence of the partial human Man II clone revealed that the DNA sequences are 84 % identical at the DNA level and 85 % identical at the amino acid level (data not shown) . Primers were designed to a region ofthe human Man II cDNA which was distinct from the murine cDNA sequence (3/17 and 7/19 mismatches, respectively) and produced a clear 178-bp PCR product with the human Man II cDNA sequence upon amplification with human genomic DNA. Amplification with hamster genomic DNA resulted in a faint band at 178 pb that was distinguishable from the human amplification product both in quantity and by the absence ofthe HindIII and EcoRI sites characteristic of the human Man II genomic sequence. DNA from a human/hamster somatic cell hybrid panel (4) was tested for the amplification of the 178-bp human Man II fragment. Cell lines which scored positive for the human Man II PCR fragment were also all found to contain human chromosome 5 (Table I) . One of the lines (hybrid line 212) contained multiple deletions in 5q and several lines contained deletions in the 5p15.1-5pl5.2 region . All of these Moremen and Robbins cDNA Cloning and Expression of Golgi a-Mannosidase 11 1531 lines scored positive for amplification of the human Man II fragment. All other chromosomes were excluded as the site for the Man II gene by discordancies in at least 15 of the hybrid lines.