Molecular cloning and expression characterization of translationally controlled tumor protein in silkworm pupae.
A Bombyx mori (B. mori) cDNA was isolated from silkworm pupae cDNA library encoding a homologue of translationally controlled tumor protein (BmTCTPk). BmTCTPk was expressed in E. coli; SDS-PAGE and Western blot showed the molecular weight of recombinant and native BmTCTPk is approximately 28 and 25 kDa, respectively; they are larger than the theoretical molecular weight. Immunohistochemical studies showed that BmTCTPk is uniformly distributed throughout the cytoplasm of BmN cells. In silkworm pupae, BmTCTPk is expressed in the midgut wall, the midgut cavity, and some fat body tissues lying between the midgut wall and body wall. Western blot and ELISAs performed on total protein extracts isolated from silkworm pupae at different development stages showed that, although BmTCTPk is expressed during all pupae stages, its expression level increases dramatically during late pupae stages, suggesting that BmTCTPk may play an important role during the developmental transition from pupa to imago.
Nie
Zuoming
Z
Institute of Biochemistry, Zhejiang Sci-Tech University, Xiasha High-Tech Zone, No. 2 Road, 310018 Hangzhou, People's Republic of China.
Lv
Zhengbing
Z
Qian
Jiawei
J
Chen
Jian
J
Li
Shanshan
S
Sheng
Qing
Q
Wang
Dan
D
Shen
Hongdan
H
Yu
Wei
W
Wu
Xiangfu
X
Zhang
Yaozhou
Y
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D015153:Blotting, Western; D012831:Bombyx / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D019295:Computational Biology; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D049750:Genome, Insect / Q000235:genetics; D007106:Immune Sera; D007150:Immunohistochemistry; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D009928:Organ Specificity; D011387:Prokaryotic Cells / Q000378:metabolism; D014176:Protein Biosynthesis / Q000235:genetics*; D021381:Protein Transport; D011679:Pupa / Q000201:enzymology / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
D018076:DNA, Complementary; D007106:Immune Sera; D019476:Insect Proteins; D009363:Neoplasm Proteins; D011994:Recombinant Proteins