|
KMID : 0578319930030020189
|
|
Molecules and Cells
1993 Volume.3 No. 2 p.189 ~ p.194
|
|
|
Isolation and Molecular Characterization of a cDNA Clone for Phytolacca americana Antiviral Protein and Its Expression in Escherichia coli
|
|
Moon, Young Ho
Jeon, Hong Seob/Choi, Kyu Whan/Lee, Jong Seob
|
|
|
Abstract
|
|
|
|
In order to isolate cDNA clones encoding Phytolacca americana antiviral protein (PAP), a cDNA library was constructed in XZAP vector with poly(A)+ RNAs purified from leaves of Phytolacca americana. The cDNA library was screened with anti-PAP antibody raised in rabbit. Among the cDNA clones obtained, clone 3 carried a cDNA fragment of about 1.2 kb in size and was subjected to DNA sequence determination. The nucleotide sequence of the PAP cDNA contained an open reading frame encoding PAP of 313 amino acids. The amino acid sequence contained 22 amino acids at the N-terminus which may function as a signal sequence in transport of this protein to extracellular matrix. The mature protein exhibited the conserved region found at other ribosome-inactivating proteins (RIPs). Southern hybridization indicated that PAPS are encoded by a small multigene family present in the genome. Northern analysis with leaf poly(A)+ RNAs revealed that PAP mRNA is about 1.4 kb in size. In order to investigate the effect of PAP on the growth of E eoh, the cDNA was manipulated so as to contain the region coding for the mature protein only by polymerise chin reaction and cloned into a microbial expression vector. E coli cells expressing PAP did not grow well as compared with cells carrying the vector only. It suggests that PAP inhibits growth of E. coli presumably by inactivating ribosomes similarly to other RIPs.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|