Molecular studies on streptomyces griseus trypsin gene = 스트렙토마이세스 그리세우스 트립신 유전자에 대한 분자생물학적 연구

Cited 0 time in webofscience Cited 0 time in scopus
  • Hit : 204
  • Download : 0
Streptomyces griseus trypsin (EC 3.4.21.4) is one of the major extracellular proteinases, which is secreted by S. griseus. The coding gene of S. griseus trypsin (sprT) of S. griseus ATCC 10137 was cloned into the pUC 18. Transformants containing sprT were isolated from genomic library by using a synthetic oligonucleotide probe and characterized by direct gel hybridization and demonstrated a proteolytic activity in S. lividans 66. The intact proteinase gene could be delimited to a 1.8 kbp Bgl I fragment. The sodium dodecyl sulfate polyacrylamide gel electrophoreses of extracellular proteins produced by S. lividans 66 harboring pSGT, which is a pUJ718-2 derivative containing a 1.8 kbp Bgl I fragment, showed that the protein band responsible for the proteinase activity was located at the same position of the S. griseus trypsin purified from a pronase. Deduced amino acid sequence from the nucleotide sequence suggests that S. griseus trypsin is produced as a precursor, consising of three portions; an amino-terminal pre sequence (32 amino acid residues), a pro sequence (4 residues), and mature trypsin. The S. griseus trypsin cosists of 223 amino acids with a computed molecular weight of 23,112. The G+C content of the sprT if 72\%. The promoter-like sequence and a putative ribosomal binding site are followed by the coding sequence of S. griseus trypsin. Following a stop codon (TGA), a structure reminiscent of the Escherichia coli rho-independent transcriptional terminator exests. The predicted amino acid sequence of S. griseus trypsin differs from the published amino acid sequence [Olafson et al.(1975) Biochemistry 14, 1168-1177]. It should be corrected by the insertion of two serine residues near position 76(Ser) according to the numbering of $\alpha$-chymotypsin. The existence of the proline residues at pro and mature junction suggests that the processing of S. griseus trypsin is non-autocatalytic. This is the first report of the nucleotide sequence for a microbial ...
Advisors
Byun, Si-Myung변시명
Description
한국과학기술원 : 생물공학과,
Publisher
한국과학기술원
Issue Date
1992
Identifier
60450/325007 / 000785054
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 생물공학과, 1992.8, [ xi, 163 p. ]

URI
http://hdl.handle.net/10203/27231
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=60450&flag=dissertation
Appears in Collection
BS-Theses_Ph.D.(박사논문)
Files in This Item
There are no files associated with this item.

qr_code

  • mendeley

    citeulike


rss_1.0 rss_2.0 atom_1.0