Worm Breeder's Gazette 11(2): 56

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Isolation and Characterization of unc-104 cDNA Clones

Anthony Otsuka

Figure 1

Although the unc-104 fragments isolated from an Arhinger-Kimble 
library were instrumental in obtaining approximately 4 kb of the 6 kb 
mRNA sequence, it was desirable to obtain full length cDNA clones to 
both complete the cDNA sequence and to produce the recombinant protein 
product in bacteria.  The discrepancy between the abundance of unc-104 
cDNA clones in the Arhinger-Kimble library (approximately 1 per 10,000)
and in the Kim-Horvitz (none in 200,000) and Thacker-Capecci 
libraries (none in 100,000), led us to screen the Barstead-Waterston 
and Schauer-Wood libraries.  The abundance of cDNA clones in these 
additional libraries suggests that an insufficient number of clones 
was screened in the cases of the Kim and Thacker libraries.
cDNA clones were isolated from 2.6 million phage (1.4 million phage 
screened from the Barstead-Waterston bank, 0.6 million the Schauer-
Wood early embryonic and him-8 embryonic libraries).  The probe was 
the 5'-most cDNA fragment obtained from the library.  The results are 
shown 
below:
[See Figure 1]
The two cDNA clones from the Barstead library overlap and cover a 
region of 4.7 kb.  DNA sequencing of the cDNA clones is currently in 
progress.

Figure 1