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Title 

Molecular cloning of a cDNA encoding ribosome inactivating protein from Amaranthus viridis and its expression in E. coli

Authors 

Suk Yoon KwonChung Sun AnJang Ryol LiuSang Soo KwakHeang Soon LeeJeong Kook KimKyung Hee Paek

Publisher 

Springer Verlag (Germany)

Issue Date 

2000

Citation 

Molecules and Cells, vol. 10, no. 1, pp. 8-12

Keywords 

amaranthus viridiscDNAexpressionribosome inactivating protein

Abstract 

In order to isolate a cDNA clone of ribosome inactivating protein (RIP), a cDNA library was constructed in Uni-ZAP XL vector with poly(A) RNA purified from leaves of Amaranthus viridis. To get the probe for screening the library, PCR of phage DNA was conducted using the vector primer and degenerate primer designed from a conserved putative active site of the RIPs. Twenty-six cDNA clones from about 600,000 plaques were isolated, and one of these clones was fully sequenced. It was 1,047 bp and contained an open reading frame encoding 270 amino acids. The deduced amino acid sequence had a putative signal sequence of 17 amino acids and a putative active site (AIQMVAEAARFFKYIE) conserved in other RIPs. E. coli cells expressing A. viridis RIP cDNA did not grow well as compared to control cells, indicating that recombinant A. viridis RIP presumably inactivated E. coli ribosomes. In addition, recombinant A. viridis RIP cDNA produced by E. coli had translation inhibition activity in vitro.

ISSN 

1016-8478

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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