Variant &lt;i&gt;GDF9&lt;/i&gt; mRNA is likely not the main cause of larger litter size in Iranian Lori-Bakhtyari, Shal, Ghezel, and Afshari sheep breeds

Abstract. This study was carried out to screen the GDF9 gene and evaluate the polymorphism effect on litter size of four Iranian sheep breeds using the PCR-RFLP and PCR-SSCP methods. First, sequencing of the GDF9 gene in 16 twin-birth, 4 triplet-birth, and 2 infertile ewes showed that, in addition to G2, G3, G4, G5, and G6 mutations that have been previously reported in other breeds, a new G0 mutation, called C25T, exists in the GDF9 sequence of 1 out of 22 ewes and causes L9F substitution in the signal peptide region. None of the triplet-birth or infertile ewes carried G1, G4, G7, FecGE, G8, or FecGT mutations. In the second experiment, a large dataset was used: 605 individuals including 496 ewes (145 Afshari, 54 Shal, 126 Ghezel, and 171 Lori-Bakhtyari sheep), and 109 rams (26 Afshari, 23 Shal, 10 Ghezel, and 50 Lori-Bakhtyari sheep. There were no sheep carrying the G7, G8, or Thoka mutations. Among all 109 rams that were used in this study, none of them were homozygous for the G1 mutation. Moreover, abundance of heterozygote rams (G1/G+) varied from 0.0 (Afshari) to 28.6 % (Lori-Bakhtyari and Ghezel). The highest and the lowest frequencies of the G4 mutation were 30.6 and 3.0 % in Shal and Afshari breeds, respectively. Moreover, G4 abundance varied from 0.0 to 42.3 %, from 3.0 to 26.9, and from 3.0 to 30.6 % in rams, ewes, and overall, respectively. There was a significant difference in the abundance of G1 and G4 mutations between breeds. However, neither the G1 nor the G4 mutation was associated with litter size in Afshari, Ghezel, Lori-Bakhtyari, or Shal sheep breeds. In conclusion, the results of this study showed that GDF9 G1 and G4 mutations are not the reason for higher litter size in Iranian sheep. Moreover, the GDF9 G0 and G6 mutations do not cause triplet births or infertility in Iranian ewes. Therefore, it is unlikely that variant GDF9 mRNA induces larger litter size or infertility in Iranian ewes.

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Genetic polymorphism in fecundity gene (FecG) among Indiansheep breeds Balangir, Shahabadi and Bonpala

Indian Journal of Animal Research ◽

10.18805/ijar.9648 ◽

2016 ◽

Author(s):

Jowel Debnath ◽

Ran Vir Singh

Keyword(s):

Litter Size ◽

Transforming Growth Factor ◽

Natural Habitat ◽

Factor B ◽

Sheep Breeds ◽

Homozygous Genotype ◽

Average Litter Size ◽

Pcr Rflp ◽

Selection Of ◽

Gdf9 Gene

FecG (GDF9) is a member of the transforming growth factor-b (TGF- b) superfamily, have been shown to be essential for follicular growth and ovulation. Different mutations in FecG gene caused increased ovulation and infertility in sheep. The present study was designed for screening polymorphism of FecG gene in 250 selected ewes from different sheep flocks representing Balangir (100), Shahabadi (100) and Bonpala (50) by employing forced PCR-RFLP technique. Genomic DNA was extracted from the blood of Balangir, Shahabadi and Bonpala matured ewes with average litter size varying from 1.00± 0.00 to 1.14±0.02 at different parities. Digestion of FecG (GDF9) gene with DdeI restriction enzyme resulted into FecGHH homozygous genotype. In all three sheep breeds, genotypic frequencies of FecGHH were 100% and gene frequency of H allele was unity. This indicates that the FecG gene is fixed in the Balangir, Shahabadi and Bonpala population in the natural habitat. Litter size of Balangir and Bonpala sheep breeds were single but in Shahabadi sheep twin was recorded. In present study all the animals of three breeds were homozygous for FecG and there was no infertility observed in above mentioned breeds in field condition and organized farm, which is not in consonance with previous report. The observed effects could be caused by linkage disequilibrium with other nearby loci. The study revealed that FecG gene is not a reliable genetic marker for selection of high prolificacy in sheep.

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Evidence of positive selection at signal peptide region of interferon gamma

Bioscience Biotechnology and Biochemistry ◽

10.1080/09168451.2014.896732 ◽

2014 ◽

Vol 78 (4) ◽

pp. 588-592 ◽

Cited By ~ 4

Author(s):

Zhengshi Wang ◽

Ming Zhong ◽

Maobin Fu ◽

Tonghai Dou ◽

Zhengqian Bian

Keyword(s):

Positive Selection ◽

Interferon Gamma ◽

Signal Peptide ◽

Signal Peptide Region ◽

Peptide Region

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A controllable expression-secretion vector constructed from the multiple trp promoter-operator, the signal peptide region of the ompF gene and the trpR gene in Escherichia coli.

Agricultural and Biological Chemistry ◽

10.1271/bbb1961.50.1295 ◽

1986 ◽

Vol 50 (5) ◽

pp. 1295-1302 ◽

Cited By ~ 2

Author(s):

Masao ITOH ◽

Hirofumi AIBA ◽

Kaoru INOKUCHI ◽

Takeshi MIZUNO ◽

Kenji NAGAHARI ◽

...

Keyword(s):

Escherichia Coli ◽

Signal Peptide ◽

Signal Peptide Region ◽

Secretion Vector ◽

Trp Promoter ◽

Peptide Region

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Non-HFE mutations in haemochromatosis in China: combination of heterozygous mutations involving HJV signal peptide variants

Journal of Medical Genetics ◽

10.1136/jmedgenet-2018-105348 ◽

2018 ◽

Vol 55 (10) ◽

pp. 650-660 ◽

Cited By ~ 8

Author(s):

Tingxia Lv ◽

Wei Zhang ◽

Anjian Xu ◽

Yanmeng Li ◽

Donghu Zhou ◽

...

Keyword(s):

Iron Overload ◽

Exome Sequencing ◽

Signal Peptide ◽

Chinese Patients ◽

Aetiological Factor ◽

Compound Heterozygous ◽

Common Mutation ◽

Smad Pathway ◽

Signal Peptide Region ◽

Peptide Region

IntroductionHereditary haemochromatosis (HH) caused by a homozygous p.C282Y mutation in haemochromatosis (HFE) gene has been well documented. However, less is known about the causative non-HFE mutation. We aimed to assess mutation patterns of haemochromatosis-related genes in Chinese patients with primary iron overload.MethodsPatients were preanalysed for mutations in the classic HH-related genes: HFE, HJV, HAMP, TFR2 and SLC40A1. Whole exome sequencing was conducted for cases with variants in HJV signal peptide region. Representative variants were analysed for biological function.ResultsNone of the cases analysed harboured the HFE p.C282Y; however, 21 of 22 primary iron-overload cases harboured at least one non-synonymous variant in the non-HFE genes. Specifically, p.E3D or p.Q6H variants in the HJV signal peptide region were identified in nine cases (40.9%). In two of three probands with the HJV p.E3D, exome sequencing identified accompanying variants in BMP/SMAD pathway genes, including TMPRSS6 p.T331M and BMP4 p.R269Q, and interestingly, SUGP2 p.R639Q was identified in all the three cases. Pedigree analysis showed a similar pattern of combination of heterozygous mutations in cases with HJV p.E3D or p.Q6H, with SUGP2 p.R639Q or HJV p.C321X being common mutation. In vitro siRNA interference of SUGP2 showed a novel role of downregulating the BMP/SMAD pathway. Site-directed mutagenesis of HJV p.Q6H/p.C321X in cell lines resulted in loss of membrane localisation of mutant HJV, and downregulation of p-SMAD1/5 and HAMP.ConclusionCompound heterozygous mutations of HJV or combined heterozygous mutations of BMP/SMAD pathway genes, marked by HJV variants in the signal peptide region, may represent a novel aetiological factor for HH.

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Expression of a Tranosema rostrale polydnavirus gene in the spruce budworm, Choristoneura fumiferana

Microbiology ◽

10.1099/0022-1317-81-7-1871 ◽

2000 ◽

Vol 81 (7) ◽

pp. 1871-1880 ◽

Cited By ~ 37

Author(s):

Catherine Béliveau ◽

Marlène Laforge ◽

Michel Cusson ◽

Guy Bellemare

Keyword(s):

Signal Peptide ◽

Fat Body ◽

Choristoneura Fumiferana ◽

Northern Analysis ◽

Juvenile Hormone Esterase ◽

Signal Peptide Region ◽

Putative Signal Peptide ◽

Cysteine Rich Protein ◽

Cellular Immune ◽

Peptide Region

The endoparasitic wasp Tranosema rostrale (Ichneumonidae) transmits a polydnavirus (PDV) to its host, Choristoneura fumiferana, during oviposition. Unlike most other PDVs examined, the virus of T. rostrale (TrPDV) does not appear to play an important role in suppressing the host cellular immune response. However, it inhibits host metamorphosis. In the present study, TrPDV gene expression was examined in parasitized and virus-injected last-instar caterpillars. Northern analysis with viral DNA as a probe revealed only one detectable mRNA, of about 650 bp. The corresponding cDNA, termed TrV1, was cloned and sequenced and found to encode a protein of 103 amino acids which, following cleavage of the putative signal peptide, has a predicted molecular mass of 9·3 kDa. This protein displays limited similarity to the VHv1.4 cysteine-rich protein from the PDV of Campoletis sonorensis, mostly within the signal peptide region. By using a TrV1-specific probe, the TrV1 gene was localized to segment G of the TrPDV genome. The cuticle and fat body were identified as the principal sites of TrV1 transcription, with little transcription observed in haemocytes and midgut. Western analysis of proteins extracted from selected tissues of parasitized insects suggested that the TrV1 protein is secreted in the haemolymph. As observed for other PDVs, injection of TrPDV did not suppress transcription of the gene that encodes juvenile hormone esterase, the activity of which is inhibited by the virus. We speculate that the TrV1 protein may play a role in the inhibition of C. fumiferana metamorphosis.

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