[Cloning, expression, and alternative splicing of the novel isoform of hTCP11 gene].
OBJECTIVE
To identify a novel isoform of hTCP11 gene and investigate its expression and alternative splicing.


METHODS
According to the sequence of human ESTs which are highly homologous to hTCP11a, primers for PCR were synthesized. Then, the amplified fragments were cloned and sequenced; some methods including BLAST, ClustalW and RT-PCR were used for genomic analysis, study of alternative splicing and gene expression among multiple tissues and different testis tissues.


RESULTS
A novel isoform of hTCP11 gene was isolated. It encodes a 440 amino acid protein that is highly homologous to the mouse 566 amino acid protein which is important to sperm function because it encodes the receptor for fertilization promoting peptide (FPP). Among TCP11a, TCP11b and TCP11c, the complicated alternative splicing was found. RT-PCR analysis of RNA extracted from human tissues revealed that the gene is only expressed in fertile adult testes, but not in azoospermic patient testes, fetal testes or other human tissues.


CONCLUSIONS
Our results along with the mouse Tcp-11 function suggest that the isoforms of TCP11 gene play important roles in sperm function and fertility.
Ma
Yong-xin
YX
Department of Medical Genetics, West China Hospital, Sichuan University, Key Laboratory of Biotherapy, Ministry of Education, Chengdu, China.
Zhang
Si-zhong
SZ
Wu
Qia-qing
QQ
Sun
Yan
Y
Qiu
Wei-min
WM
Xu
Wen-ming
WM
D000328:Adult; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008297:Male; D008565:Membrane Proteins; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics*; D020033:Protein Isoforms; D017385:Sequence Homology; D056427:t-Complex Genome Region
D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D020033:Protein Isoforms; C533399:TCP11 protein, human; C533400:Tcp11 protein, mouse