pMIB/V5-His is a 3.6 kb vector designed for constitutive expression and secretion of recombinant proteins from Sf9, Sf21, and High Five insect cells. Secretion of proteins into serum-free medium simplifies purification, making it easier to harvest protein from cultured cells. pMIB/V5-His has several features to facilitate expression of recombinant proteins in insect cells including:
• The OpIE2 promoter for constitutive expression
• The honeybee melittin (HBM) secretion signal to allow secreted expression of your protein
• The Blasticidin resistance gene for rapid selection of stably transfected cell lines in two weeks
• C-terminal V5-epitope and polyhistidine tag for detection with Invitrogens Anti-V5 antibody and purification with nickel-chelating resin
引用及参考文献
Antibodies against a secreted protein from hookworm larvae reduce the intensity of hookworm infection in humans and vaccinated laboratory animals.
Bethony J, Loukas A, Smout M, Brooker S, Mendez S, Plieskatt J, Goud G, Bottazzi ME, Zhan B, Wang Y, Williamson A, Lustigman S, Correa-Oliveira R, Xiao S, Hotez PJ, FASEB J (2005) 19:1743-1745
Product usage: ASP-2 was fused in-frame into pMIB-V5/HisA using the SphI and XbaI restriction sites. Protein was expressed in Spodpotera frugiperda Sf9 insect cells.
CATGATGATAAACAATGTATGGTGCTAATGTTGCTTCAACAACAATTCTGTTGAACTGTGTTTTCATGTT
TGCCAACAAGCACCTTTATACTCGGTGGCCTCCCCACCACCAACTTTTTTGCACTGCAAAAAAACACGCT
TTTGCACGCGGGCCCATACATAGTACAAACTCTACGTTTCGTAGACTATTTTACATAAATAGTCTACACC
GTTGTATACGCTCCAAATACACTACCACACATTGAACCTTTTTGCAGTGCAAAAAAGTACGTGTCGGCAG
TCACGTAGGCCGGCCTTATCGGGTCGCGTCCTGTCACGTACGAATCACATTATCGGACCGGACGAGTGTT
GTCTTATCGTGACAGGACGCCAGCTTCCTGTGTTGCTAACCGCAGCCGGACGCAACTCCTTATCGGAACA
GGACGCGCCTCCATATCAGCCGCGCGTTATCTCATGCGCGTGACCGGACACGAGGCGCCCGTCCCGCTTA
TCGCGCCTATAAATACAGCCCGCAACGATCTGGTAAACACAGTTGAACAGCATCTGTTCGAATTTAAAGC
TACCATGAAATTCTTAGTCAACGTTGCCCTTGTTTTTATGGTCGTATACATTTCTTACATCTATGCCGGC
ATGCTAAGCTTGGTACCGAGCTCGGATCCACTAGTCCAGTGTGGTGGAATTCTGCAGATATCCAGCACAG
TGGCGGCCGCTCGAGGTCACCCATTCGAAGGTAAGCCTATCCCTAACCCTCTCCTCGGTCTCGATTCTAC
GCGTACCGGTCATCATCACCATCACCATTGAGTTTATCTGACTAAATCTTAGTTTGTATTGTCATGTTTT
AATACAATATGTTATGTTTAAATATGTTTTTAATAAATTTTATAAAATAATTTCAACTTTTATTGTAACA
ACATTGTCCATTTACACACTCCTTTCAAGCGCGTGGGATCGATGCTCACTCAAAGGCGGTAATACGGTTA
TCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTA
AAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTC
AAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTG
CGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGC
TTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCA
CGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGA
CACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTA
CAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCT
GAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGT
GGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTT
CTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGCGAAACACGCACGG
CGCGCGCACGCAGCTTAGCACAAACGCGTCGTTGCACGCGCCCACCGCTAACCGCAGGCCAATCGGTCGG
CCGGCCTCATATCCGCTCACCAGCCGCGTCCTATCGGGCGCGGCTTCCGCGCCCATTTTGAATAAATAAA
CGATAACGCCGTTGGTGGCGTGAGGCATGTAAAAGGTTACATCATTATCTTGTTCGCCATCCGGTTGGTA
TAAATAGACGTTCATGTTGGTTTTTGTTTCAGTTGCAAGTTGGCTGCGGCGCGCGCAGCACCTTTGCCGG
GATCTGCCGGGCTGCAGCACGTGTTGACAATTAATCATCGGCATAGTATATCGGCATAGTATAATACGAC
AAGGTGAGGAACTAAACCATGGCCAAGCCTTTGTCTCAAGAAGAATCCACCCTCATTGAAAGAGCAACGG
CTACAATCAACAGCATCCCCATCTCTGAAGACTACAGCGTCGCCAGCGCAGCTCTCTCTAGCGACGGCCG
CATCTTCACTGGTGTCAATGTATATCATTTTACTGGGGGACCTTGCGCAGAACTCGTGGTGCTGGGCACT
GCTGCTGCTGCGGCAGCTGGCAACCTGACTTGTATCGTCGCGATCGGAAATGAGAACAGGGGCATCTTGA
GCCCCTGCGGACGGTGCCGACAGGTTCTTCTCGATCTGCATCCTGGGATCAAAGCCATAGTGAAGGACAG
TGATGGACAGCCGACGGCAGTTGGGATTCGTGAATTGCTGCCCTCTGGTTATGTGTGGGAGGGCTAAGCA
CTTCGTGGCCGAGGAGCAGGACTGACACGTCCCGGGAGATCTGCATGTCTACTAAACTCACAAATTAGAG
CTTCAATTTAATTATATCAGTTATTACCCATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTC
GCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAA
AAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCT
TGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTA
TTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTG
AGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCAT
AACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCT
TTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATAC
CAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGA
ACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTT
CTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCG
GTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCA
GGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTG
TCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGG
TGAAGATCCTTTTTGATAATCT