Abstract:
【Objective】 To describe the molecular characteristics of
SPOP and
MyD88 genes and their expression in different tissue during chicken development, and lay the basis for the subsequent study of their regulation of tissue growth and development in chickens and the development of disease-resistance breeding.【Method】 The coding regions(CDS) of chicken
SPOP and
MyD88 genes were cloned by RT-PCR. ExPASy, SOPMA, SWISS-MODEL and PSORT II Prediction were used for bioinformatics analysis. Real-time quantitative PCR(qRT-PCR) technology was used to detect the expression of
SPOP and
MyD88 genes in various tissues at 14 embryonic days(E14 d) in chick embryos and at 1-day-old(H1 d), 7-day-old(H7 d), and 14-day-old(H14 d) after shelling.【Result】 The CDS sequences of chicken
SPOP and
MyD88 genes were 1125 and 900 bp, encoding 374 and 299 amino acid residues, respectively. The molecular formula of the
SPOP protein was C
1866H
2926N
496O
559S
28, and was predicted to have a molecular weight of 42 kD and a isoelectric point(pI) of 5.58. The molecular formula of the
MyD88 protein was C
1502H
2394N
410O
438S
18 and was predicted to have a molecular weight of 33 kD and a pI of 5.93. Both
SPOP and
MyD88 were relatively unstable proteins. The chicken
SPOP and
MyD88 proteins, were mainly located in the cytoplasm(60.9%) and their secondary structures mainly consisted of α -helixes and random loops. Compared with humans and mammals, the three functional domains of chicken SPOP(MATH, BTB-POz and BACK) were conserved, while the two functional domains of MyD88(Death and TIR) had multiple amino acid site variations.
SPOP and
MyD88 genes were expressed in all tissues of chicken at different developmental stages, but their expression in the lung was relatively the highest. The differences in expression of
SPOP and
MyD88 was extremely significant(
P<0.01, the same below). From E14 d to H14 d, the expression of
SPOP gene in the eyeball and lung showed an overall upward trend, but the lung expression of
SPOP stabilized before H14 d. The expression of
SPOP gene in the brain, heart, stomach muscle and liver showed an upward trend followed by a downward trend. The expression of
MyD88 gene in the eyeball, liver and lung was first increased, followed by a decrease and a further increase. The expression of
MyD88 gene in stomach and chest muscles decreased and reached the lowest value at H14 d.【Conclusion】 The expression levels of
SPOP and
MyD88 genes are relatively high in lung, eyeball and stomach muscle of chicken embryo at different developmental stages. The expression levels of
SPOP are extremely significantly higher than
MyD88 and the relative expression levels of
SPOP gene and
MyD88 gene are negatively correlated, indicating that
SPOP gene regulates the expression of
MyD88 gene.