| RRC ID |
541
|
| Author |
Funaguma S, Shimada T.
|
| Title |
Accurate pre-mRNA splicing using a nuclear extract from Bombyx mori fat body.
|
| Journal |
Insect Biochem Mol Biol
|
| 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.
|
| Volume |
35(3)
|
| Pages |
257-61
|
| Published |
2005-3-1
|
| DOI |
10.1016/j.ibmb.2004.12.005
|
| 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
|
| Times Cited |
0
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
ENTOMOLOGY
|
| Resource |
| Silkworms |
|
