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. 1987 Nov;102(5):1289-301.
doi: 10.1093/oxfordjournals.jbchem.a122167.

In vitro splicing of a chicken delta-crystallin pre-mRNA in a mammalian nuclear extract

H Sakamoto  1 , M Ohno, K Yasuda, K Mizumoto, Y Shimura
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Free article

In vitro splicing of a chicken delta-crystallin pre-mRNA in a mammalian nuclear extract

H Sakamoto et al. J Biochem. 1987 Nov.
Free article

Abstract

An in vitro splicing system was constructed using portions of chicken delta-crystallin pre-mRNA synthesized in vitro and a HeLa nuclear extract. Analysis of the reaction products revealed that about 25% of the pre-mRNA was precisely spliced at 30 degrees C in 2 h under the standard conditions. The other major products of the reaction detected were a 5'-exon fragment and three RNA species showing unusual electrophoretic mobilities on polyacrylamide gels. Structural analyses showed that these three RNAs contain a branch (lariat) structure as seen in the in vitro splicing reactions of human beta-globin, adenovirus, and yeast pre-mRNAs. In addition, methylation at the N-7 position of the blocking guanosine of the 5'-terminal cap structure of pre-mRNA has been suggested to play an important role in the splicing reaction.

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