1. 1. Conditions for in vitro translation of mRNA in cell-free extracts from cultured Spodoptera frugiperda cells were defined. 2. 2. Incorporation of [35S]methionine into acid-precipitable material increased for approximately 1 hr, and was sensitive to the protein synthesis inhibitors pactamycin and cycloheximide. 3. 3. Micrococcal nuclease-treated lysate, primed with purified rabbit globin mRNA, synthesized a major protein with the size of full length globin, indicating that the lysate supported correct initiation and elongation of polypeptides.
|Original language||English (US)|
|Number of pages||4|
|Journal||Comparative Biochemistry and Physiology -- Part B: Biochemistry and|
|State||Published - 1989|
Bibliographical noteFunding Information:
translationfr om an exogenoums RNA, extractw as treatedw ith micrococcanl ucleasea ccordingt o the procedured escribedb y Gillies and Stollar (1981)t o reducet he amounto f endogenoums RNA, and the treatede xtracwt ass upplementewdit hp urifiedg lobin mRNA. Analysiso f the translatiopnr oductso n an SDS gel showeda substantiallrye ducedi ncorpora-tion of \[3S5 \]methionininet o labelledp olypeptideisn the treated,g lobin-supplementeexdt ract,w ith the Acknowledgements--This work was supportedb y Grant AI 20385f rom the National Institutesa t Health, by GrantS O7 RR05908a dministerebdy the NIH Divisiono f ResearchR esourcest hrought he Universityo f Medicine andD entistroy f N.J. Schoolo f OsteopathMice dicinea, nd by the Universityo f MinnesotaA griculturaEl xperiment StationT. his is contributio1n6 ,538fr omthe Universitoy f MinnesotaE xperimenStt ation,S t Paul, MN. We thank LethaF ieldsf or typingt hem anuscripatn, dD r T. J. Kurtti for readingth emanuscript.