Cloning and characterization of testis-specific tektin in Bombyx mori.
A testis-specific cDNA library of Bombyx mori was constructed by an mRNA subtraction technique. Several clones were randomly selected and determined for their nucleotide sequences. One of them, designated as BmTST, contained a 3'-part of an open reading frame homologous to tektin, the protein known to form filamentous polymers in the walls of ciliary and flagellar microtubules. Also isolated was a genomic fragment, which contains the 5'-part of the coding sequence of BmTST and its promoter region. As a whole, the complete open reading frame was found to encode 508 amino acid residues, whose sequence had 28, 28 and 30% identities with the Strongylocentrotus purpuratus tektins A1, B1 and C1, respectively. Expression analysis by reverse transcription polymerase chain reaction with the cDNA and Western blotting with a polyclonal antibody indicated that the BmTST gene was expressed specifically in the testis during sperm maturation. The protein was immunologically detected exclusively in the fraction expected to contain the 9 + 2 flagellar axonemes of sperms. We infer that the BmTst protein is possibly involved in the spermatogenesis of B. mori.
Ota
Atsuko
A
Laboratory of Sericultural Science, Faculty of Agriculture, Kyushu University Graduate School, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan.
Kusakabe
Takahiro
T
Sugimoto
Yasushi
Y
Takahashi
Masateru
M
Nakajima
Yumiko
Y
Kawaguchi
Yutaka
Y
Koga
Katsumi
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D012831:Bombyx / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling; D015723:Gene Library; D008297:Male; D008868:Microtubule Proteins / Q000032:analysis / Q000737:chemistry* / Q000235:genetics*; D008870:Microtubules / Q000737:chemistry; D008969:Molecular Sequence Data; D009928:Organ Specificity; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013094:Spermatozoa / Q000737:chemistry; D013737:Testis / Q000166:cytology / Q000378:metabolism*
D018076:DNA, Complementary; D008868:Microtubule Proteins; D012333:RNA, Messenger; C044103:tektins