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求救引物设计

丁香园论坛

1360
1: AF075241. Sus scrofa prepro...[gi:3328338] Links

LOCUS AF075241 451 bp mRNA linear MAM 17-MAY-2000
DEFINITION Sus scrofa prepro-orexin mRNA, complete cds.
ACCESSION AF075241
VERSION AF075241.1 GI:3328338
KEYWORDS .
SOURCE Sus scrofa (pig)
ORGANISM Sus scrofa
Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi;
Mammalia; Eutheria; Cetartiodactyla; Suina; Suidae; Sus.
REFERENCE 1 (bases 1 to 451)
AUTHORS Dyer,C.J., Touchette,K.J., Carroll,J.A., Allee,G.L. and
Matteri,R.L.
TITLE Cloning of porcine prepro-orexin cDNA and effects of an
intramuscular injection of synthetic porcine orexin-B on feed
intake in young pigs
JOURNAL Domest. Anim. Endocrinol. 16 (3), 145-148 (1999)
MEDLINE 99275384
PUBMED 10343916
REFERENCE 2 (bases 1 to 451)
AUTHORS Matteri,R.L. and Dyer,C.J.
TITLE Direct Submission
JOURNAL Submitted (29-JUN-1998) Agricultural Research Service, USDA, Room
S-143 ASRC, University of Missouri, Columbia, MO 65211, USA
FEATURES Location/Qualifiers
source 1..451
/organism="Sus scrofa"
/mol_type="mRNA"
/db_xref="taxon:9823"
/tissue_type="hypothalamus"
/dev_stage="neonate"
CDS 1..396
/function="neuropeptide precursor; involved in appetite
control"
/codon_start=1
/product="prepro-orexin"
/protein_id="AAC26827.1"
/db_xref="GI:3328339"
/translation="MNPPFAKVSWATVTLLLLLLLLPPAVLSPGAAAQPLPDCCRQKT
CSCRLYELLHGAGNHAAGILTLGKRRPGPPGLQGRLQRLLQASGNHAAGILTMGRRAG
AEPAPRLCPGRRCLAAAASSVAPGGRSGI"
sig_peptide 1..99
mat_peptide 100..198
/product="orexin-A"
mat_peptide 208..291
/product="orexin-B"
misc_feature 1..393
/note="encodes prepro-orexin"
misc_feature 100..393
/note="encodes pro-orexin
1 atgaatcctc cttttgcaaa ggtctcctgg gccaccgtga cgctgctgct tctgctactg
61 ttgctgccgc ctgcggtgct gtcgcccggg gcggcggcgc agccactgcc cgactgctgc
121 cgtcaaaaga cgtgctcctg ccgcctctac gagctgctac acggcgctgg caatcacgcg
181 gccggcatcc tcactctggg caagcggcgg ccagggcccc cgggcctgca aggccggctg
241 cagcgcctcc tgcaggccag cggcaaccac gcagcgggca tcctgaccat gggccgccgc
301 gcaggcgcag agccagcgcc gcgcctctgc ccggggcgcc ggtgtctcgc tgcggctgcc
361 tcatctgtag cgccgggagg acggtctggg atctgagccc tagcccggcc ctctcccctc
421 tgcccgcccg aggtcagccc cccagaaaaa g

本人查阅资料有人做过用他的引物却怎么也扩增不出来,扩增出来的不是我的目的片段大小,我的目的片段大小为356,可扩增出来的主要为二百多的片段。正义引物5‘-CGCTGCTGCTTCTGCTACTG-3’反义引物5‘-TCAGATCCCAGACCGTCCTC-3’
请各位高手帮帮,我急用。
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