Cloning and sequencing of GH, H-FABP and HSP72 genes in yak

2007 ◽  
Vol 87 (4) ◽  
pp. 503-510
Author(s):  
Jin-cheng Zhong ◽  
Zhi-jie Ma ◽  
Jiang-tao Ou ◽  
Xiao Zhu ◽  
Sheng-gang Liu

The growth hormone (GH) gene, the heart fatty acid binding protein (H-FABP) gene and the heat shock protein 72 (HSP72) gene of the yak (Bos grunniens) were cloned and sequenced. The sequences were compared with those of other animals, and phylogenetic trees were constructed by the NJ (neighborhood joining) method. Results showed that the yak GH gene was composed of five exons (13, 161, 117, 162 and 198 bp) and four introns (248, 225, 229 and 275 bp). The cDNA of GH was a 654 bp nucleotide encoding a putative protein of 217 amino acid (AA) residuals, with a signal peptide of 26 AAs and the mature peptide of 191 AAs. The H-FABP gene was composed of four exons (73, 173, 102 and 54 bp) and three introns (3460, 1892 and 1495 bp). The cDNA was 402 bp encoding a putative protein of 133 AAs. The yak HSP72 gene was an intron-free 1926bp nucleotide, encoding a protein of 641 AAs. The data suggest that the three genes from the yak were highly conserved with other species at the nucleic acid level. The results of the phylogenetic trees reflect the molecular evolution relationship among these species, consistent with the zoological classification. The genetic distance calculated by the nucleotide sequence of the GH gene’s coding region indicated that genetic distances among yak, cattle, gayal and zebu were relatively small, but genetic distances between these four species and buffalo were relatively large. Therefore, it was more reasonable to consider yak, cattle, gayal and zebu as independent species of the genus Bos, while buffalo belongs to another category (Bubalus). Key words: Yak, GH gene, H-FABP gene, HSP72 gene, phylogenetic evolution

Author(s):  
Hui Wang ◽  
Hanbo Zhao ◽  
Yujia Chu ◽  
Jiang Feng ◽  
Keping Sun

Abstract High-frequency hearing is particularly important for echolocating bats and toothed whales. Previously, studies of the hearing-related genes Prestin, KCNQ4, and TMC1 documented that adaptive evolution of high-frequency hearing has taken place in echolocating bats and toothed whales. In this study, we present two additional candidate hearing-related genes, Shh and SK2, that may also have contributed to the evolution of echolocation in mammals. Shh is a member of the vertebrate Hedgehog gene family and is required in the specification of the mammalian cochlea. SK2 is expressed in both inner and outer hair cells, and it plays an important role in the auditory system. The coding region sequences of Shh and SK2 were obtained from a wide range of mammals with and without echolocating ability. The topologies of phylogenetic trees constructed using Shh and SK2 were different; however, multiple molecular evolutionary analyses showed that those two genes experienced different selective pressures in echolocating bats and toothed whales compared to non-echolocating mammals. In addition, several nominally significant positively selected sites were detected in the non-functional domain of the SK2 gene, indicating that different selective pressures were acting on different parts of the SK2 gene. This study has expanded our knowledge of the adaptive evolution of high-frequency hearing in echolocating mammals.


1996 ◽  
Vol 319 (2) ◽  
pp. 483-487 ◽  
Author(s):  
Claire MEUNIER-DURMORT ◽  
Hélène POIRIER ◽  
Isabelle NIOT ◽  
Claude FOREST ◽  
Philippe BESNARD

The role of fatty acids in the expression of the gene for liver fatty acid-binding protein (L-FABP) was investigated in the well-differentiated FAO rat hepatoma cell line. Cells were maintained in serum-free medium containing 40 µM BSA/320 µM oleate. Western blot analysis showed that oleate triggered an approx. 4-fold increase in the cytosolic L-FABP level in 16 h. Oleate specifically stimulated L-FABP mRNA in time-dependent and dose-dependent manners with a maximum 7-fold increase at 16 h in FAO cells. Preincubation of FAO cells with cycloheximide prevented the oleate-mediated induction of L-FABP mRNA, showing that protein synthesis was required for the action of fatty acids. Run-on transcription assays demonstrated that the control of L-FABP gene expression by oleate was, at least in part, transcriptional. Palmitic acid, oleic acid, linoleic acid, linolenic acid and arachidonic acid were similarly potent whereas octanoic acid was inefficient. This regulation was also found in normal hepatocytes. Therefore long-chain fatty acids are strong inducers of L-FABP gene expression. FAO cells constitute a useful tool for studying the underlying mechanism of fatty acid action.


2008 ◽  
Vol 60 (2) ◽  
pp. 408-413 ◽  
Author(s):  
F.C. Figueiredo ◽  
P.S. Lopes ◽  
A.P.G. Pinto ◽  
D.A.F. Paiva ◽  
P.T. Mendonça ◽  
...  

The heart fatty acid-binding protein (HFABP) gene was sequenced in parental animals of a F2 crossing of boars of the Brazilian native Piau breed with commercial sows (Landrace x Large White Pietrain). Primers used for PCR were designed to amplify four exons of the gene. The PCR products were sequenced and compared with the GenBank sequences. Differences between the generated sequences and the GenBank sequences were observed for both genetic groups. A total of 246 F2 animals were genotyped using the Hinf I restriction enzyme. Two genotypes were identified, 198 being animals HH and 48 Hh. The Hinf I SNP was significantly associated with weights of loin (bone-in) (P<0.05), jowl (P<0.05), sirloin (P<0.10), and kidneys (P<0.01). These results showed the potential of the H-FABP gene in marker-assisted selection programs for carcass traits in pigs.


2019 ◽  
Vol 5 (Supplement_1) ◽  
Author(s):  
Monika Olech ◽  
Jacek Kuźmak

Abstract Previous phylogenetic analyses of small ruminant lentivirus (SRLV) sequences found in Poland revealed the circulation of subtype A1 in both sheep and goats, subtypes B1 in goats, and subtypes B2, A12, and A13 in sheep only. This study aimed to analyze the genetic nature of SRLV circulating in sheep and goats from single-species flocks. In order to analyze the degree of genetic variability, the fragments of gag and env genes of 24 SRLV strains were amplified by PCR, cloned into plasmid vectors, sequenced, and consensus sequences were aligned to each other and to reference sequences available from GenBank. Phylogenetic analysis was performed using the Geneious tree-builder tool, and phylogenetic trees were constructed using Mr Bayes (using the general time reversible substitution model) within Geneious Pro 5.3. Pairwise genetic distances were calculated in MEGA 6. Phylogenetic analysis revealed that the strains were highly heterogeneous and represented ovine strains belonging to subtypes A12 and B2 and caprine strains grouped in subtypes B1, B2, A1, and A12. In addition, two novel subtypes, A16 and A17, were found in goats. The mean pairwise genetic distances of gag and env sequences of both clusters were above 15 per cent nucleotide divergence when compared to all other subtypes within group A, which is a criterion required to distinguish a new subtype. Additionally, the existence of two separated clusters was confirmed by high bootstrap values. Co-infections with strains belonging to different subtypes within A and B groups were detected in one sheep and four goats originating from four flocks. Since the co-infection with more than one lentivirus genotype offers an opportunity for viral recombination, the possible recombination events were tested based on RDP analysis. For all co-infected animals, no evidence of recombination was found within the gag gene; however, env sequences showed some recombination patterns in three samples. In conclusion, we have demonstrated extended genetic variability of SRLV in sheep and goats from Poland with the existence of co-infection and recombination events.


2019 ◽  
Vol 20 (23) ◽  
pp. 5948 ◽  
Author(s):  
Wang ◽  
Yue ◽  
Liu ◽  
Yang ◽  
Li ◽  
...  

The fatty acid-binding protein (FABP) gene family, which encodes a group of fatty acid-trafficking molecules that affect cellular functions, has been studied extensively in mammals. However, little is known about the gene structure, expression profile, and regulatory mechanism of the gene family in chickens. In the present study, bioinformatics-based methods were used to identify the family members and investigate their evolutionary history and features of gene structure. Real-time PCR combined with in vivo and in vitro experiments were used to examine the spatiotemporal expression pattern, and explore the regulatory mechanism of FABP genes. The results show that nine members of the FABP gene family, which branched into two clusters and shared a conserved FATTYACIDBP domain, exist in the genome of chickens. Of these, seven FABP genes, including FABP1, FABP3-7, and FABP10 were abundantly expressed in the liver of hens. The expression levels of FABP1, FABP3, and FABP10 were significantly increased, FABP5 and FABP7 were significantly decreased, and FABP4 and FABP6 remained unchanged in hens at the peak laying stage in comparison to those at the pre-laying stage. Transcription of FABP1 and FABP3 were activated by estrogen via estrogen receptor (ER) α, whilst FABP10 was activated by estrogen via ERβ. Meanwhile, the expression of FABP1 was regulated by peroxisome proliferator activated receptor (PPAR) isoforms, of which tested PPARα and PPARβ agonists significantly inhibited the expression of FABP1, while tested PPARγ agonists significantly increased the expression of FABP1, but downregulated it when the concentration of the PPARγ agonist reached 100 nM. The expression of FABP3 was upregulated via tested PPARβ and PPARγ agonists, and the expression of FABP7 was selectively promoted via PPARγ. The expression of FABP10 was activated by all of the three tested PPAR agonists, but the expression of FABP4-6 was not affected by any of the PPAR agonists. In conclusion, members of the FABP gene family in chickens shared similar functional domains, gene structures, and evolutionary histories with mammalian species, but exhibited varying expression profiles and regulatory mechanisms. The results provide a valuable resource for better understanding the biological functions of individual FABP genes in chickens.


1990 ◽  
Vol 110 (5) ◽  
pp. 1791-1801 ◽  
Author(s):  
K A Roth ◽  
J M Hertz ◽  
J I Gordon

The gastrointestinal tract is lined with a monolayer of cells that undergo perpetual and rapid renewal. Four principal, terminally differentiated cell types populate the monolayer, enterocytes, goblet cells, Paneth cells, and enteroendocrine cells. This epithelium exhibits complex patterns of regional differentiation, both from crypt-to-villus and from duodenum-to-colon. The "liver" fatty acid binding protein (L-FABP) gene represents a useful model for analyzing the molecular basis for intestinal epithelial differentiation since it exhibits cell-specific, region-specific, as well as developmental stage specific expression. We have previously linked portions of the 5' nontranscribed domain of the rat L-FABP gene to the human growth hormone (hGH) gene and analyzed expression of the fusion gene in adult transgenic mice. High levels of hGH expression were noted in enterocytes as well as cells that histologically resembled enteroendocrine cells. In the present study, we have used immunocytochemical techniques to map the distribution of enteroendocrine cells in the normal adult mouse gut and to characterize those that synthesize L-FABP. In addition, L-FABP/hGH fusion genes were used to identify subsets of enteroendocrine cells based on their ability to support hGH synthesis in several different pedigrees of transgenic mice. The results reveal remarkable differences in transgene expression between, and within, enteroendocrine cell populations previously classified only on the basis of their neuroendocrine products. In some cases, these differences are related to the position occupied by cells along the duodenal-to-colonic and crypt-to-villus axes of the gut. Thus, transgenes appear to be sensitive tools for examining the cellular and regional differentiation of this class of intestinal epithelial cells.


2011 ◽  
Vol 91 (4) ◽  
pp. 593-596 ◽  
Author(s):  
Jun He ◽  
Lizhi Lu ◽  
Yong Tian ◽  
Zhengrong Tao ◽  
Deqian Wang ◽  
...  

He, J., Lu, L., Tian, Y., Tao, Z., Wang, D., Li, J., Li, G., Shen, J., Fu, Y. and Niu, D. 2011. Short Communication: Analysis of intramuscular fat and fatty acids of different duck breeds and their association with SNPs of duck A-FABP gene. Can. J. Anim. Sci. 91: 593–596. Intramuscular fat (IMF) is related to organoleptic characteristics of meat. Adipocyte fatty acid-binding protein (A-FABP) is one of the intracellular lipid-binding proteins involved in the transportation of fatty acids. The IMF contents of six duck breeds were measured, and the complete sequence and part of the 5' flanking region of duck A-FABP gene were obtained in this study. The IMF contents of different breeds were significantly different (P<0.05). Two SNPs were detected in the exon 3, one (HQ640428: g.2018A>G) was significantly associated with the contents of three fatty acids, total IMF and pectoral muscle weight. This work provides useful data for duck breeding.


ISRN Agronomy ◽  
2012 ◽  
Vol 2012 ◽  
pp. 1-11 ◽  
Author(s):  
Chi Zhang ◽  
Arun S. Pratap ◽  
S. Natarajan ◽  
L. Pugalendhi ◽  
Shinji Kikuchi ◽  
...  

Cucumber, Cucumis sativus (), and melon, Cucumis melo (), are two common vegetable crops worldwide. The present study evaluated eighteen Cucumis accessions (nine C. sativus and nine C. melo) that were collected from three South Asian countries that have the most diversity of Cucumis. Nine quantitative and twenty-three qualitative characteristics were measured. The values of fruit weight displayed the biggest divergence among the nine quantitative traits and much variation was displayed in twenty-three qualitative traits among eighteen accessions. For eight morphological quantitative traits other than fruit weight, eighteen accessions were divided into three groups by using Principle Component Analysis and K-means cluster analysis. Also, two chloroplast genes rbcL and matK of eighteen accessions were sequenced. Combined sequences were subjected to construct phylogenetic trees by Neighbor-Joining and Maximum Likelihood methods. Topologies of nine melon accessions were same in these two methods and nine cucumber accessions showed difference. The genetic distances among each of C. sativus and C. melo accessions were not high. We conclude that the genetic relationship among the eighteen accessions used in this study is not distant although they display significant morphological variations. The information on novel Cucumis germplasms provided here would contribute to breeding program as well as evolutional study in Cucumis.


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