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Chinese Journal of Oceanology and Limnology 2013年第1期31卷 118-127页

作者：蒋克勇孙姝娟刘梅王宝杰孟晓林王雷 R&D Center of Marine Biotechnology Institute of OceanologyChinese Academy of Sciences Graduate University of Chinese Academy of Sciences

AMP deaminase catalyzes the conversion of AMP into IMP and ammonia. In the present study, a full-length cDNA of AMPD1 from skeletal muscle of Japanese flounder Paralichthys olivaceus was cloned and characterized. The 2 526 bp cDNA contains a 5'-UTR of 78 bp, a 3'-UTR of 237 bp and an open reading frame (ORF) of 2 211 bp, which encodes a protein of 736 amino acids. The predicted protein contains a highly conserved AMP deaminase motif (SLSTDDP) and an ATP-binding site sequence (EPLMEEYAIAAQVFK). Phylogenetic analysis showed that the AMPD1 and AMPD3 genes originate from the same branch, but are evolutionarily distant from the AMPD2 gene. RT-PCR showed that the flounder AMPD1 gene was expressed only in skeletal muscle. QRT-PCR analysis revealed a statistically significant 2.54 fold higher level of AMPD1 mrna in adult muscle (750±40 g) compared with juvenile muscle (7.5±2 g) (Pexpression level and IMP content in the skeletal muscle of juvenile and adult flounders. These results may provide useful information for quality improvement and molecular breeding of aquatic animals.

关键词： AMPD1 cDNA cloning mrna expression IMP skeletal muscle Japanese flounder

cDNA cloning and mrna expression of the translationally controlled tumor protein(TCTP)gene from Japanese sea perch(Lateolabrax japonicus)

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Acta Oceanologica Sinica 2005年第2期24卷 113-119页

作者： QIULihua SONGLinsheng WULongtao XUWei JIANGShigui InstituteofOceanology ChineseAcademyofSciencesQingdao266071China／／SouthChinaSeaFisheriesResearchInstituteChineseAcademyofFisherySciencesGuangzhou510300China InstituteofOceanology ChineseAcademyofSciencesQingdao266071China InstituteofOceanology ChineseAcademyofSciencesQingdao266071China／／GraduateSchoolChineseAcademyofSciencesBeijingI00039China SouthChinaSeaFisheriesResearchInstitute ChineseAcademyofFisherySciencesGuangzhou510300China

A homologue of the lower vertebrates translationally controlled tumor protein (TCTP) was cloned from the marine fish Japanese sea perch (Lateolabrax japonicus) by the technology of homology cloning. The full-length cDNA sequence of the sea perch TCTP gene contained a 5' untranslated region (UTR) of 47 bp, a 3' UTR of 433 bp, and a putative open reading frame (ORF) of 510 bp encoding a polypeptide of 170 amino acids. The deduced amino acid sequence of the sea perch TCTP gene showed a high similarity to that of zebrafish, rohu, rabbit, chicken and human. Sequence analysis revealed there were a signature sequence of TCTP family, an N-glycosylation site, and five Casein kinase phosphorylation sites in the sea perch TCTP. The temporal expression of TCTP genes in healthy and lipopolysaccharide (LPS) challenged fishes was measured by semi-quantitative reverse transcription-PCR (RT-PCR). The results indicated that LPS could up-regulate the expression of sea perch TCTP in the examined tissues, including head-kidney, spleen and liver.

关键词： translationally controlled tumor protein Lateolabrax japonicus cDNA cloning mrna expression

Effects of Dietary Lysine on Growth Performance, Serum Concentrations of Insulin-Like Growth Factor-I (IGF-I) and IGF-I mrna expression in Growing Rabbits

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Agricultural Sciences in China 2010年第6期9卷 887-895页

作者： JING Wen-qian LI Fu-chang College of Animal Science and Technology Shandong Agricultural University Tai＇an 271018 P.R.China College of Life Science Linyi Normal University Linyi 276005 P.R.China

The effects of dietary lysine on production performance,serum concentrations of metabolites,growth hormone （GH）,insulin-like growth factor-I （IGF-I） and IGF-I mrna expression in growing rabbits were *** hundred weaned New Zealand rabbits were allocated to individual cages and randomly offered a diet containing 5.5 （L1）,6.5 （L2）,7.5 （L3）,8.5 （L4）,or 9.5 g （L5） lysine per kg *** results showed that the average daily gain （ADG） of the rabbits from L3,L4 or L5 was higher than those from L1 or L2 （P 〈 0.05）.The feed gain ratio （F/G） in the rabbits from L4 or L5 was lower than those from L1 or L2 （P 〈 0.05）.Dietary lysine did not affect serum concentrations of total protein （TP）,glucose,insulin （INS）,and growth hormone （GH）（P 〉 0.05）.The quadratic effects of lysine on the serum urea nitrogen （SUN） concentration was detected （P = 0.035）.Serum IGF-I concentrations had a trend to increase quadratically with the increasing dietary lysine （P = 0.07）.A significant correlation was found between serum IGF-I concentrations （x,ng mL-1） and ADG （y,g kg-1）： y = -0.017x2 ＋ 1.984x ＋ 20.87 （R2 = 0.8982,P = 0.003）.The relative abundance of hepatic and muscular IGF-I mrna tended to increase with increasing dietary lysine levels （P = 0.053 and 0.082,respectively）.Providing the diets mainly consisted of corn,wheat bran and peanut vine,the most appropriate dietary lysine level for growing meat rabbits from weaning to 70 d old was found to be 8.5 g kg-1,and IGF-I may be an important factor controlling growth of weaned rabbits.

关键词： New Zealand rabbit lysine growth performance IGF-I mrna expression

Differential mrna expression levels and gene sequences of carboxylesterase in both deltamethrin resistant and susceptible strains of the cotton aphid, Aphis gossypii

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Insect Science 2008年第3期15卷 209-216页

作者： Chuan-Wang Cao Jing Zhang Xi-Wu Gao Pei Liang Hui-Lin Guo Department of Entomology China Agricultural University Beijing China

Extensive use of insecticides on cotton has prompted resistance development in the cotton aphid, Aphis gossypii （Glover） in China, A deltamethrin-selected population of cotton aphids from Xinjiang Uygur Autonomous Region, China with 228,59-fold higher resistance to deltamethrin was used to examine how carboxylesterase conferred resistance to this pyrethroid insecticide. The carboxylesterase activity in the deltamethrin-resistant strain was 3.67-, 2,02- and 1.16-fold of the susceptible strain when using α-naphthyl acetate （α-NA）, β-naphthyl acetate （β-NA） and α-naphthyl butyrate （α-NB） as substrates, respectively, Carboxylesterase cDNA was cloned and sequenced from both deltamethrinresistant and susceptible strains. The cDNA contained 1581 bp open reading frames （ORFs） coding a 526 amino acid protein. Only one amino acid substitution （Val^87-Ala） was observed between deltamethrin-resistant and susceptible strains but it is not genetically linked to resistance by the catalytic triad and signature motif analysis. The real-time polymerase chain reaction analysis indicated that the resistant strain had a 6.61-fold higher level of carboxylesterase mrna than the susceptible strain. The results revealed that up-regulation of the carboxylesterase gene, not modified gene structure, may be responsible for the development of resistance in cotton aphids to deltamethrin.

关键词： Aphis gossypii (Glover) carboxylesterase deltamethrin resistance genesequence mrna expression

Cloning and mrna expression of macrophage migration inhibitory factor (MIF) gene of large yellow croaker (P seudosciaena crocea)

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Acta Oceanologica Sinica 2010年第3期29卷 63-73页

作者： MAO Yong XU Bing SU Yongquan ZHANG Zhiwen DING Shaoxiong WANG Ding WANG Jun Fisheries College of Guangdong Ocean University Zhanjiang 524025 China State Key Laboratory of Marine Environmental Science Xiamen University Xiamen 361005 China

Mammalian macrophage migration inhibitory factor （MIF） plays an important role as an indispensable mediator in the pathogenesis of inflammatory disease like septicemia, but little is known about the role of MIF homologue in fish septicemia. The authors have cloned the MIF homologue in large yellow croaker Pseudosciaena crocea （LycMIF） using RACE approach. The full-length cDNA of LycMIF was 634 bases and contained an ORF of 345 bases encoding a protein of 115 amino acid residues. As demonstrated by RT-PCR and QRT-PCR assay, MIF mrnas were constitutively expressed in 11 selected tissues and were abundant in brain and liver. Moreover, the LycMIF transcripts in the liver and head kidney were responsive to bacteria infection and could be significantly up-regulated. Our results provide the first direct evidence that fish MIF was implicated in pathogenesis of fish vibrosis and play an important role in response to bacteria infection.

关键词： macrophage migration inhibitory factor (MIF) vibriosis large yellow croaker mrna expression

The mrna expression and Methylation Status in Imprinting Control Region of H19 Gene Between Cattle-Yak and Their Parents

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Journal of Integrative Agriculture 2012年第10期11卷 1691-1699页

作者： LI Ming-gui LIU Zhen-shan PAN Zeng-xiang LUO Hua XIE Zhuang LI Qi-fa College of Animal Science and Technology Nanjing Agricultural University

The H19 gene, which is imprinted with preferential expression from the maternal allele, was one of the first identified imprinting genes in mammals. Recent studies revealed that correct imprinting of the H19 gene plays a vital role in human spermatogenesis. To investigate whether imprinting defects were associated with the hybrid sterility of male cattle-yak, the methylation patterns of the H19 imprinting control region （ICR） and H19 mrna expression in the testes of cattle-yak, yak, and cattle were examined. The results showed that the 3rd CCCTC-binding factor （CTCF） site of the H19 ICR was significantly hypomethylated in the testes of cattle-yak compared with yak or cattle. As expected, H19 was expressed at a significantly higher level in cattle-yak than in yak or cattle. These results suggest that imprinting defects of the CTCF- binding site in the HI9 ICR were possibly associated with disturbed spermatogenesis of male cattle-yak. Thus, we propose that disorders in H19 imprinting, resulting in an increased H19 mrna expression, might contribute to the sterility of F1 male hybrids between cattle and yak.

关键词： cattle-yak male sterility H19 ICR CTCF-binding site mrna expression DNA methylation

Effects of supplements differing in fatty acid profile to late gestational beef cows on cow performance,calf growth performance,and mrna expression of genes associated with myogenesis and adipogenesis

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Journal of Animal Science and Biotechnology 2021年第4期12卷 1483-1497页

作者： Taoqi Shao Frank A.Ireland Joshua C.McCann Daniel W.Shike Department of Animal Sciences University of Illinois at Urbana-ChampaignUrbanaILUSA

Background:Maternal nutrition during gestation affects fetal development,which has long-term programming effects on offspring postnatal growth *** a critical role in protein and lipid metabolism,essential fatty acids can influence the development of muscle and adipose *** experiment investigated the effects of late gestation supplements(77 d prepartum),either rich in saturated and monounsaturated fatty acids(CON;155 g/cow/d EnerGII)or polyunsaturated fatty acids(PUFA;80 g/cow/d Strata and 80 g/cow/d Prequel),on cow performance and subsequent calf growth performance as well as mrna expression in longissimus muscle(LM)and subcutaneous adipose tissue at birth and ***:There was no difference(P≥0.34)in cow body weight(BW)or body condition score from presupplementation through *** concentrations of C18:3n-3 and C20:4n-6 decreased(P≤0.05)to a greater extent from mid-supplementation to calving for PUFA compared with CON *** plasma C20:0,C20:5n-3,and C22:6n-3 were increased(P≤0.01)in PUFA during supplementation *** birth,PUFA steers had greater(P=0.01)plasma C20:*** differences(P≥0.33)were detected in steer birth BW or dam milk production,however,CON steers tended(P=0.06)to have greater pre-weaning average daily gain and had greater(P=0.05)weaning BW compared with *** mrna expression in steers:MYH7 and C/EBPβin LM increased(P≤0.04)to a greater extent from birth to weaning for PUFA compared with CON;MYF5 in LM and C/EBPβin adipose tissue tended(P≤0.08)to decrease more from birth to weaning for CON compared with PUFA;SCD in PUFA adipose tissue tended(P=0.08)to decrease to a greater extent from birth to weaning than *** addition,maternal PUFA supplementation tended(P=0.08)to decrease MYOG mrna expression in LM and decreased(P=0.02)ZFP423 in adipose tissue during the pre-weaning ***:Late gestation PUFA supplementation decreased pre-weaning growth performance of the subsequent steer progeny compared

关键词： Beef cattle Fatty acids Fetal programming Late gestation mrna expression

Analysis of IGF2 mrna expression and its methylation status between cattle yaks and their parents

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Progress in Natural Science:Materials International 2009年第9期19卷 1063-1069页

作者： Zhenshan Liu a,b,1, Qifa Li a,1, Qingbo Zhang a, Xuguang Qu a, Liyan Dong a, Chamba Yang-zom c, Zhuang Xie a,*, Honglin Liu a a Laboratory of Animal Reproductive Genetics and Molecular Evolution, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China b Institute of Animal Husbandry and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan 250100, China c College of Agriculture and Animal Husbandry, Tibet University, Linzhi 860000, China Laboratory of Animal Reproductive Genetics and Molecular Evolution College of Animal Science and Technology Nanjing Agricultural University Nanjing 210095 China Institute of Animal Husbandry and Veterinary Medicine Shandong Academy of Agricultural Sciences Jinan 250100 China College of Agriculture and Animal Husbandry Tibet University Linzhi 860000 China

Cattle yaks are an F1 hybrid between cattle and yaks and exhibit significant hybrid vigor. However, cattle yaks are sterile males. To study the epigenetic aspects of this reproductive isolation, IGF2 mrna expression in cattle yaks and their parents and the patterns of the IGF2 differentially methylated regions (DMR) were examined in blood and testes. The results showed that IGF2 expression in cattle yaks’ testes was lower than in their parents (P 0.05). Our study showed that IGF2 plays an important role in bovine spermato-genesis and might be involved in cattle-yak male sterility. The methylation level of the IGF2 DMR was irrelevant to the lower expression of IGF2 in cattle yaks; other factors perhaps play roles in its expression.

关键词： Cattle yak Male sterility IGF2 gene mrna expression DNA methylation

Progastrin-releasing peptide and gastrin-releasing peptide receptor mrna expression in non-tumor tissues of the human gastrointestinal tract

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World Journal of Gastroenterology 2006年第16期12卷 2574-2578页

作者： Hans-Jürg Monstein Niclas Grahn Mikael Truedsson Bodil Ohisson Molecular Biology Laboratory-LMC University HospitalDepartment of Biomedicine and SurgeryFaculty of Health SciencesS-581 85 LinkpingSweden Department of Clinical Sciences University Hospital MalmS-205 02 MalmSweden

AIM： To investigate the expression of gastrin-releasing peptide （GRP） and GRP-receptor mrna in non-tumor tissues of the human esophagus, gastrointestinal tract, pancreas and gallbladder using molecular biology techniques. METHODS： Poly A^＋ mrna was isolated from total RNA extracts using an automated nucleic acid extractor and, subsequently, converted into single-stranded cDNA （sscDNA）. PCR amplifications were carried out using genespecific GRP and GRP-receptor primers. The specificity of the PCR amplicons was further confirmed by Southern blot analyses using gene-specific GRP and GRP-receptor hybridization probes. RESULTS： expression of GRP and GRP-receptor mrna was detected at various levels in nearly all segments of the non-tumor specimens analysed, except the gallbladder. In most of the biopsy specimens, coexpression of both GRP and GRP-receptor mrna appeared to take place. However, expression of GRP mrna was more prominent than was GRP-receptor mrna. CONCLUSION： GRP and GRP-receptor mrnas are expressed throughout the gastrointestinal tract and provides information for the future mapping and determination of its physiological importance in normal and tumor cells.

关键词： Gastrin releasing peptide (GRP) Gastrinreleasing peptide receptor (GRPR) mrna expression Morphogenesis Gastrointestinal tract

Single Nucleotide Polymorphism of CYP3A4 Intron 2 and Its Influence on CYP3A4 mrna expression and Liver Enzymatic Activity in Human Liver

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2015年第4期35卷 502-507页

作者：黄敏王汉明郭瑜平洁陈曼徐丹汪晖 Department of Endocrinology Hubei Integrative Chinese and Western Medicine Hospital Department of Pharmacology Basic Medical School of Wuhan University Hubei Provincial Key Laboratory of Developmentally Originated Disease

Summary： In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of en- dogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism （SNP） of CYP3A4 intron 2 in the liver and its effects on the mrna expression and enzymatic activity of CYP3A4, genomic DNA was extracted from 96 liver tissue samples obtained from patients who had undergone liver surgery. An SNP of CYP3A4 intron 2 was identified by polymerase chain reaction （PCR）-single-strand confirmation polymorphism and DNA sequencing. The mrna expression of CYP3A4 was determined by the fluorescence quantitative PCR technique. The enzymatic activity of CYP3A4 was measured using erythromycin and testosterone as probe substrates. Twelve patients were found to have the SNP/T4127G CYP3A4 within intron 2. The mrna levels of CYP3A4 in wild-type and SNP/T4127G samples were 2.62±1.09 and 2.79±1.63, respectively （P〉0.05）. Erythromycin N-demethylase activity in wild-type and SNP/T4127G samples were 121.2±32.8 and 124.7±61.6 nmol·mg^-1min^-1, respectively （P〉0.05）. The activity of testosterone 613-hydroxylase was significantly different between wild-type （648±173 pmol·mg^-1·min^-1） and SNP/T4127G samples （540-4-196 ***-l-minl; P〈0.05）. In conclusion, the SNP/T4127G of CYP3A4 intron 2 exists in the liver. This SNP does not affect the mrna expression of CYP3A4 but significantly decreases the hepatic micro- somal testosterone 613-hydroxylase activity of CYP3A4. Furthermore, this study indicates that the ap- propriate selection of probe substrates is very important in studying the relationship between the geno- type and phenotype of CYP3A4.

关键词： CYP3A4 single nucleotide polymorphism mrna expression enzymatic activity human liver