123 

 end (444 bases sequenced from the T3 promoter) was 

 translated to yield a sequence having about 23 to 25% 

 identity with the DQT functional domain of the 

 pentafunctional amino acid sequence from S. cerevisiae or A. 

 nidulans, respectively, and about 22% identity with the 

 monofunctional AroD sequence from E. coli, Fig. 7-2. The 

 sequence determination at the 3' end (sequenced from the T7 

 promoter) was not informative at this stage because the 

 deduced amino acid sequence showed very little conservation 

 between the known genes of E. coli, S. cerevisiae or A. 

 nidulans at the C-terminal region. 



Partial sequencing of 5' and 3' ends of several of the 

 possible ADH and PAT cDNA clones showed no significant 

 identity to known genes. 

 Subcloninq cDNA encoding the S -protein 



Since the partial sequence at the 5 'end of the cloned 

 insert, the positive functional complementation results and 

 the direct assay of activities were all mutually reinforcing 

 that the cDNA encoding the S -protein had been cloned, the 

 SP3 clone was used further for subcloning analysis in order 

 to obtain a complete nucleotide sequence. 



Based on the multiple cloning sites of the vectors to 

 be used for subcloning, pUC18, pGEMSZf (+) and pBluescript+, 

 restriction enzymes were chosen and tested in order to 

 locate and map the restriction sites present in the SP3 cDNA 

 clone. Analysis of agarose gels (Fig. 7-lB, right and 



"■'-^ 



