[cDNA cloning and mRNA expression pattern in follicles of the mature inhibin alpha subunit from Xianju chicken].
The mature region of Xianju chicken inhibin alpha-subunit was amplified from the total RNA of follicle granulosa cells by RT-PCR using the primer pair designed according to the reported cDNA sequence of chicken inhibin alpha-subunit, and this fragment of alpha-subunit was cloned and sequenced subsequently. The results revealed that the mature alpha-subunit of Xianju chicken was a fragment of 113 amino acids containing one glycosylation site and seven cysteine residues. It was approximately 98% and 61.4%-68.7% identical in nucleotide sequence, 97.3% and 64.6%-69% similar in deduced amino acid sequence, respectively, in the mature region to the chicken and mammalian inhibin alpha-subunit cDNA clone. As for the mature chicken inhibin alpha-subunit, the number of potential glycosylation site and cysteine residues was the same, and their corresponding positions in the amino acid sequences were almost identical as compared to chicken and mammalian inhibin alpha-subunit, which indicated that the inhibin alpha-subunit was highly conserved among different species, implying an important role of inhibin alpha-subunit in various animals. The quantitative analysis of competitive RT-PCR for inhibin alpha-subunit revealed that the expression of alpha-subunit decreased with further follicle maturity from SYF to F1 follicle. The highest level of inhibin alpha-subunit mRNA was found in the SYF and F6-8 follicles, which indicated that inhibin alpha-subunit played an important role during the course of follicular recruitment, selection and dominance.
Fu
Y
Y
Animal Science College of Zhejiang University, Hangzhou 310029, China.
Niu
D
D
Ruan
H
H
Yu
X P
XP
Chen
G
G
He
G Q
GQ
Yang
P X
PX
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002645:Chickens / Q000235:genetics*; D003001:Cloning, Molecular; D005260:Female; D006031:Glycosylation; D007265:Inhibins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D006080:Ovarian Follicle / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; C094377:inhibin-alpha subunit; D007265:Inhibins