Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (6): 279-290.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1152
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WANG Nan(), ZHANG Rui, PAN Yang-yang, HE Hong-hong, WANG Jing-lei, CUI Yan, YU Si-jiu()
Received:
2021-09-07
Online:
2022-06-26
Published:
2022-07-11
Contact:
YU Si-jiu
E-mail:594302635@qq.com;sjyu@163.com
WANG Nan, ZHANG Rui, PAN Yang-yang, HE Hong-hong, WANG Jing-lei, CUI Yan, YU Si-jiu. Cloning of Bos grunniens TGF-β1 Gene and Its Expression in Major Organs of Female Reproductive System[J]. Biotechnology Bulletin, 2022, 38(6): 279-290.
引物Primer | 引物序列Primer sequence(5'-3') | 产物长度Product size/bp | 退火温度Annealing temperature Tm/℃ | GenBank登录号GenBank No. |
---|---|---|---|---|
D-TGF-β1 | F:ACTACTACGCCAAGGAGGTCAC | 153 | 60 | MZ004937 |
R:TTCCGAGAATCTTAGCTGCA | ||||
TGF-β1 | F:GCAAACAGACCCTCCTACCTTT | 1 173 | 60 | MZ004937 |
R:TCCTTAAATACAGTCCTGGTGAG | ||||
GAPDH | F:GGGTCATCATCTCTGCACCT | 178 | 60 | EU195062.1 |
R:TGGTCATAAGTCCCTCCACG |
Table 1 Primer sequences and length
引物Primer | 引物序列Primer sequence(5'-3') | 产物长度Product size/bp | 退火温度Annealing temperature Tm/℃ | GenBank登录号GenBank No. |
---|---|---|---|---|
D-TGF-β1 | F:ACTACTACGCCAAGGAGGTCAC | 153 | 60 | MZ004937 |
R:TTCCGAGAATCTTAGCTGCA | ||||
TGF-β1 | F:GCAAACAGACCCTCCTACCTTT | 1 173 | 60 | MZ004937 |
R:TCCTTAAATACAGTCCTGGTGAG | ||||
GAPDH | F:GGGTCATCATCTCTGCACCT | 178 | 60 | EU195062.1 |
R:TGGTCATAAGTCCCTCCACG |
Fig. 1 D-TGF-β1 and GAPDH PCR amplification electrop-horesis M:DL2000 DNA marker. 1-3:Fallopian tubes,uterus and ovaries in luteal phase. 4-6:Fallopian tubes,uterus and ovaries in pregnancy phase. 7-8:Fallopian tubes,uterus and ovaries in follicular phase
Fig. 5 Results of yak TGF-β1 protein via bioinformatics analysis A:The secondary structure prediction of TGF-β1 protein in Bos grunniens. The long vertical zone:Alpha helix(h). The middle vertical zone:Extended stand(e). The short vertical zone:Random coil(c). B:The tertiary structure prediction of TGF-β1 protein in Bos grunniens. C:Hydrophilicity and hydrophobicity of the protein encoded by TGF-β1 gene. D:The protein transmembrane regional analysis of TGF-β1 protein in Bos grunniens. E:TGF-β1 phosphorylation site analyses
Fig. 6 Relative expressions of TGF-β1 mRNA in different tissues A:Ovary. B:Fallopian tubes. C:Uterus. Reference gene:GAPDH;n=4. Different letters indicate significant difference(P<0.05),the same below
Fig. 7 Detection results of TGF-β1 and GAPDH protein in different tissues A:Ovary. B:Fallopian tubes. C:Uterus. 1:Follicular phase. 2:Luteal phase. 3:Pregnancy
Fig. 9 Distribution of TGF-β1 proteins in different tissues A-D:Positive expression. A,A1,A2,A3,B,B1,and B2:Ovary. C,C1,and C2:Fallopian tube. D,D1,D2:Uterus. E-H:Negative control.SG:Follicle granular layer. EM:Mucosalepithelium. GE:Germinal epitnelium. TF:Theca follicle. CL:Corpus luteum verum. EP:Epitheliummucosae.UG:Uterineglands. sg:Serous gland
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