Buck (Capra hircus) genes encode new members of the spermadhesin family.
Spermadhesins are the major proteins of boar seminal plasma and form a group of polypeptides probably involved in reproduction. In previous work, a member of the spermadhesin family from buck seminal plasma, called BSFP, was characterized by mass spectrometry and N-terminal sequencing. The present study aimed to clone and characterize the BSFP gene and investigate its expression along the genital tract using real-time polymerase chain reaction (PCR). The cDNAs of the seminal vesicle, testis, epididymis, bulbourethral gland, and ductus deferens were prepared from a buck. Following 3'- and 5'-end amplifications using seminal vesicle cDNA, we cloned and sequenced four highly similar (97-98%) nucleotide sequences encoding spermadhesins, which were named Bodhesin-1(Bdh-1), Bdh-2, Bdh-3, and Bdh-4. All deduced amino acid sequences contained the CUB domain signature and were 49-52% similar to boar AWN. Among the four Bdh amino acid sequences, Bdh-2 was the most similar to the BSFP N-terminal fragment. By using real-time PCR, it was verified specific amplifications for all Bdh in the seminal vesicle, testis, epididymis, and bulbourethral gland, with the exception of Bdh-2 in epididymis. The amplicons had a melting temperature and size of approximately 78 degrees C and 130 bp, respectively. Bdh expression was higher in the seminal vesicle when compared to the other tissues. The present work confirms that goat is the fifth mammalian species, after pig, cattle, horse, and sheep, in which spermadhesin molecules are found. To the best of our knowledge, this is the first report on buck spermadhesin genes using molecular cloning and expression profile.
Melo
Luciana Magalhães
LM
Laboratório de Moléculas Biologicamente Ativas-Biomol-Lab., Department of Biochemistry and Molecular Biology, Federal University of Ceará, Avenida Humberto Monte s/n, bloco 907, sala 1075, Campus do Pici, Fortaleza-CE, Brazil.
Teixeira
Dárcio Italo Alves
DI
Havt
Alexandre
A
da Cunha
Rodrigo Maranguape Silva
RM
Martins
Danyelly Bruneska Gondim
DB
Castelletti
Carlos Henrique Madeiros
CH
de Souza
Paulo Roberto Eleutério
PR
Filho
José Luiz de Lima
JL
Freitas
Vicente José de Figueirêdo
VJ
Cavada
Benildo Sousa
BS
Rádis-Baptista
Gandhi
G
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D005837:Genitalia, Male / Q000378:metabolism*; D006041:Goats / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D029589:Seminal Plasma Proteins / Q000235:genetics*; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D029589:Seminal Plasma Proteins; C425042:spermadhesin