In:
Intervirology, S. Karger AG, Vol. 51, No. 2 ( 2008), p. 144-150
Abstract:
〈 i 〉 Objectives: 〈 /i 〉 To construct a one-plasmid expression system of the armored RNA containing long chimeric RNA byincreasing the number and affinity of the 〈 i 〉 pac site 〈 /i 〉 . 〈 i 〉 Methods: 〈 /i 〉 The plasmid pET-MS2- 〈 i 〉 pac 〈 /i 〉 was constructedwith one C-variant 〈 i 〉 pac site, 〈 /i 〉 and then the plasmid pM-CR-2C containing 1,891-bp chimeric sequences and two C-variant 〈 i 〉 pac sites 〈 /i 〉 was produced. Meanwhile, three plasmids (pM-CR-C, pM-CR-2W and pM-CR-W) were obtained as parallel controls with a different number and affinity of the 〈 i 〉 pac site 〈 /i 〉 . Finally, the armored RNA was expressed and purified. 〈 i 〉 Results: 〈 /i 〉 The armored RNA with 1,891 bases target RNA was expressed successfully by the one-plasmid expression system with two C-variant 〈 i 〉 pac sites 〈 /i 〉 , while for one 〈 i 〉 pac site 〈 /i 〉 , no matter whether the affinity was changed or not, only the 1,200 bases target RNA was packaged. It was also found that the C-variant 〈 i 〉 pac site 〈 /i 〉 could increase the expression efficiency of the armored RNA. The armored RNA with 1,891-bp exogenous RNA in our study showed the characterization of ribonuclease resistance and stability at different time points and temperature conditions. 〈 i 〉 Conclusions: 〈 /i 〉 The armored RNA with 1,891 bases exogenous RNA was constructed and the expression system can be used as a platform for preparation of the armored RNA containing long RNA sequences.
Type of Medium:
Online Resource
ISSN:
0300-5526
,
1423-0100
Language:
English
Publisher:
S. Karger AG
Publication Date:
2008
detail.hit.zdb_id:
1482863-7
SSG:
12
Permalink