Characterization of the Luteinizing Hormone Beta (LH-β) Subunit Gene in the Goat

2021 ◽  
Author(s):  
Jakeer Husen A Olekar ◽  
Ashish Kumar ◽  
Ambika Sharma ◽  
Kranti Dev
Keyword(s):  
Luteinizing Hormone ◽  
Genomic Dna ◽  
Critical Role ◽  
Glycosylation Site ◽  
Restriction Enzyme Digestion ◽  
Β Subunit ◽  
Dna Encoding ◽  
Result Analysis ◽  
Cloned Gene

Background: Luteinizing hormone (LH) plays a critical role in ovulation and maintenance of pregnancy in female and gamete production in male during fertile phase of life. Physiological disturbance of this hormone leads to conditions like delayed ovulation, anovulation, cystic ovarian disease and lack of sexual desire in male. Since, there had been no report of molecular characterization of β-subunit of luteinizing hormone (LH) of Indian goat, the present study aimed to clone and characterize genomic DNA encoding LHβ subunit.Methods: Genomic DNA was extracted from goat blood and amplified using specific LHβ gene primers. After cloning and transformation, plasmids were isolated from randomly selected white colonies. Presence of insert was confirmed by restriction enzyme digestion of plasmids. After confirmation by PCR, plasmids were sent for DNA sequencing. Result: Analysis of sequence revealed an insert of 1006 bp size as expected. Comparison of nucleotide sequence revealed the cloned gene to be is LHβ encoding 141 amino acids. It showed 97.3 and 91.7% similarity with sheep and cattle respectively. Inferred amino acid sequence showed absolute similarity (100%) with sheep and buffalo. The common and essential features such as twelve cysteine molecules, a single potential N-glycosylation site, the CAGY region and another tetrapeptide CGPC are all found in the goat sequence too.

scholarly journals Molecular cloning and characterization of pirarucu (Arapaima gigas) follicle-stimulating hormone and luteinizing hormone β-subunit cDNAs

PLoS ONE ◽  
2017 ◽  
Vol 12 (8) ◽  
pp. e0183545 ◽  
Author(s):  
Thais Sevilhano ◽  
Roberto Feitosa de Carvalho ◽  
Nélio Alessandro de Jesus Oliveira ◽  
João Ezequiel Oliveira ◽  
Vinicius Gonçalves Maltarollo ◽  
...  
Keyword(s):  
Luteinizing Hormone ◽  
Molecular Cloning ◽  
Follicle Stimulating Hormone ◽  
Β Subunit ◽  
Arapaima Gigas ◽  
Stimulating Hormone

Insect immunity: molecular cloning, expression, and characterization of cDNAs and genomic DNA encoding three isoforms of insect defensin in Aedes aegypti

1999 ◽  
Vol 8 (1) ◽  
pp. 107-118 ◽  
Author(s):  
C. A. Lowenberger ◽  
C. T. Smartt ◽  
P. Bulet ◽  
M. T. Ferdig ◽  
D. W. Severson ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Molecular Cloning ◽  
Genomic Dna ◽  
Insect Immunity ◽  
Dna Encoding ◽  
Insect Defensin

Characterization of cDNA and genomic DNA encoding the a subunit of a human GTP-binding protein Gi

1987 ◽  
Vol 19 ◽  
pp. S10-S10
Author(s):  
S KIM ◽  
S ANG ◽  
R LEE ◽  
Y KAWAHARA ◽  
J SEIDMAN ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Binding Protein ◽  
Dna Encoding ◽  
Gtp Binding Protein ◽  
Gtp Binding ◽  
A Subunit

Characterization of the luteinizing hormone beta (LH-β) subunit gene in the Indian river buffalo (Bubalus bubalis)

2008 ◽  
Vol 155 (1) ◽  
pp. 63-69 ◽  
Author(s):  
Mallikarjuna Shivapura Basavarajappa ◽  
Sachinandan De ◽  
Manish Thakur ◽  
Tirtha Kumar Datta ◽  
Gaurav Dogra ◽  
...  
Keyword(s):  
Luteinizing Hormone ◽  
Bubalus Bubalis ◽  
River Buffalo ◽  
Β Subunit ◽  
Subunit Gene ◽  
Indian River

Characterization of cDNA and Genomic DNA Encoding SERCA1, the Ca2+-ATPase of Human Fast-Twitch Skeletal Muscle Sarcoplasmic Reticulum, and Its Elimination as a Candidate Gene for Brody Disease

Genomics ◽  
1995 ◽  
Vol 30 (3) ◽  
pp. 415-424 ◽  
Author(s):  
Yilin Zhang ◽  
Junichi Fujii ◽  
Michael S. Phillips ◽  
Hai-Shiene Chen ◽  
George Karpati ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Sarcoplasmic Reticulum ◽  
Candidate Gene ◽  
Genomic Dna ◽  
Dna Encoding ◽  
Fast Twitch

scholarly journals Roles of N-linked and O-linked glycosylation sites in the activity of equine chorionic gonadotropin in cells expressing rat luteinizing hormone/chorionic gonadotropin receptor and follicle-stimulating hormone receptor

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
So-Yun Lee ◽  
Munkhzaya Byambaragchaa ◽  
Seung-Hee Choi ◽  
Han-Ju Kang ◽  
Myung-Hwa Kang ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Luteinizing Hormone ◽  
Chorionic Gonadotropin ◽  
Follicle Stimulating Hormone ◽  
Glycosylation Site ◽  
Β Subunit ◽  
Α Subunit ◽  
Post Transfection ◽  
Glycosylation Sites ◽  
Equine Chorionic Gonadotropin

Abstract Background Equine chorionic gonadotropin (eCG), which comprises highly glycosylated α-subunit and β-subunit, is a unique member of the glycoprotein hormone family as it elicits both follicle-stimulating hormone (FSH)-like and luteinizing hormone (LH)-like responses in non-equid species. To examine the biological function of glycosylated sites in eCG, the following glycosylation site mutants were constructed: eCGβ/αΔ56, substitution of Asn56 of α-subunit with Gln; eCGβ-D/α, deletion of the O-linked glycosylation site at the carboxyl-terminal peptide (CTP) region of the β-subunit; eCGβ-D/αΔ56, double mutant. The recombinant eCG (rec-eCG) mutants were expressed in Chinese hamster ovary suspension (CHO-S) cells. The FSH-like and LH-like activities of the mutants were examined using CHO-K1 cells expressing rat lutropin/CG receptor (rLH/CGR) and rat FSH receptor (rFSHR). Results Both rec-eCGβ/α and rec-eCGβ/αΔ56 were efficiently secreted into the CHO-S cell culture medium on day 1 post-transfection. However, the secretion of eCGβ-D/α and eCGβ-D/αΔ56, which lack approximately 12 O-linked glycosylation sites, was slightly delayed. The expression levels of all mutants were similar (200–250 mIU/mL) from days 3 to 7 post-transfection. The molecular weight of rec-eCGβ/α, rec-eCGβ/αΔ56 and rec-eCG β-D/α were in the ranges of 40–45, 37–42, and 34–36 kDa, respectively. Treatment with peptide-N-glycanase F markedly decreased the molecular weight to approximately 5–10 kDa. Rec-eCGβ/αΔ56 exhibited markedly downregulated LH-like activity. The signal transduction activity of both double mutants was completely impaired. This indicated that the glycosylation site at Asn56 of the α-subunit plays a pivotal role in the LH-like activity of eCG. Similarly, the FSH-like activity of the mutants was markedly downregulated. eCGβ-D/α exhibited markedly downregulated LH-like and FSH-like activities. Conclusions Rec-eCGβ/α exhibits potent biological activity in cells expressing rLH/CGR and rFSHR. The findings of this study suggest that the LH-like and FSH-like activities of eCG are regulated by the N-linked glycosylation site at Asn56 of the eCG α-subunit and/or by the O-linked glycosylation sites of the eCG β-subunit. These findings improved our understanding of the mechanisms underlying both LH-like and FSH-like activities of eCG.

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