Abstract:
:The computer program, SIGSEQ2, was used to select heterologous signal peptides from a catalog of published sequences to express the echistatin gene in insect (Sf9) cells. S-values for each amino acid were determined to select empirically the site of cleavage between the signal peptide and mature echistatin. Five gene fragments encoding the signal peptides for human immunoglobulin kappa (Ig kappa), Drosophila 68C glue, antistasin, bovine growth hormone (bGH), and human apolipoprotein E (Apo E) were constructed by the use of long synthetic oligonucleotides or polymerase chain reaction (PCR). Echistatin expression vectors then were constructed using the baculovirus polyhedrin promoter. Following transient transfection, the media were assayed for echistatin activity. The results indicate that the computer program greatly facilitated the selection and design of five different signal peptides and accurately predicted their relative functionality in the expression and secretion of echistatin in insect cell cultures.
journal_name
DNA Cell Bioljournal_title
DNA and cell biologyauthors
Daugherty BL,Zavodny SM,Lenny AB,Jacobson MA,Ellis RW,Law SW,Mark GEdoi
10.1089/dna.1990.9.453subject
Has Abstractpub_date
1990-07-01 00:00:00pages
453-9issue
6eissn
1044-5498issn
1557-7430journal_volume
9pub_type
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