RRC ID |
541
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著者 |
Funaguma S, Shimada T.
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タイトル |
Accurate pre-mRNA splicing using a nuclear extract from Bombyx mori fat body.
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ジャーナル |
Insect Biochem Mol Biol
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Abstract |
A cell-free pre-mRNA splicing system was developed from the nuclear extract of Bombyx mori fat bodies. The synthetic pre-mRNA from the cytoplasmic actin gene of B. mori (BmA3 ) was used as a substrate for in vitro splicing. The pre-mRNA was accurately spliced in the nuclear extract prepared from the fat body. Sequencing of the amplicon by RT-PCR revealed that the spliced site in vitro was identical to the site in vivo. We found that a part of the input capped pre-mRNA was accurately spliced after 2h of incubation. In addition, the RT-PCR analysis precisely mapped the branch point to an adenine located at 30 nucleotides before the 3' end of the first intron in BmA3. This system was a first example that differentiated tissue in insects yielded active extracts for in vitro splicing.
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巻・号 |
35(3)
|
ページ |
257-61
|
公開日 |
2005-3-1
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DOI |
10.1016/j.ibmb.2004.12.005
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PII |
S0965-1748(04)00208-5
|
PMID |
15705505
|
MeSH |
Actins / genetics
Animals
Bombyx / chemistry*
Cell Extracts / chemistry
Cell Nucleus / chemistry*
Fat Body / chemistry*
RNA Precursors / chemistry*
RNA Precursors / genetics
RNA Splicing*
|
IF |
3.827
|
引用数 |
0
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WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
ENTOMOLOGY
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リソース情報 |
カイコ |
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