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Neuroglobin mutation associated with hypoxia adaptation in Tibet chicken

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Progress in Natural science:Materials International 2007年第12期17卷 1419-1424页

作者： Zhang lifan1, liu Chong2, lian linsheng3, zhao chunjiang1, bao haigang1 and wu changxin1(1. college of animal science and technology, china agricultural university, beijing 100094, china 2. college of Biological science, china agricultural university, beijing 100094, china 3. college of animal science and technology, Yunnan agricultural university, Kunming 650201, china) college of animal science and technology China Agricultural University Beijing 100094 China college of Biological science China Agricultural University Beijing 100094 China college of animal science and technology Yunnan Agricultural University Kunming 650201 China

Globin protein family plays an important role in storing and transporting oxygen. As a newly reported globin, the revealed function of neuroglobin includes binding and storing oxygen as well as facilitating the utilization of oxygen in neuronal cells. In the present study, mutations in exons of chicken Ngb gene were identified with the method of sequencing and created restriction site PCR in Tibet chicken and other four lowland chicken breeds. The mutations of Lys-2224(E4)-Asn and Ser-2279(E4)-Gly were identified in exon 4 of the gene. The Lys-2224(E4)-Asn mutation existed only in Tibet chicken and the mutation frequencies increased with increasing altitude. Analysis of the haplotype and diplotype of the two mutations in Tibet chicken populations of different altitudes showed that the frequencies of TG haplotype and TTGG diplotype also increased with increasing altitude, while the reverse tendency was found on GGAA diplotype. Under the hypoxic simulation incubation, the main haplotype was TG in living embryos and GA in dead embryos. The results showed that the Lys-2224(E4)-Asn mutation may be a specific mutation associated with hypoxia adaptation in Tibet chicken.

关键词： Tibet chicken neuroglobin hypoxia adaptation Ngb gene mutation.

Reduction of drosopterin content caused by a 45-nt insertion in Henna pre-mRNA of Drosophila melanogaster

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science china(life sciences) 2008年第8期51卷 702-710页

作者： WANG Qin, zhao chunjiang, BAI liHua, DENG XueMei & wu changxin State Key Laboratory of Agrobiotechnology & Key laboratory of animal Genetics and Breeding of the Ministry of Agriculture, college of animal science and technology, china agricultural university, beijing 100193, china State Key Laboratory of Agrobiotechnology & Key laboratory of animal Genetics and Breeding of the Ministry of Agriculture College of Animal Science and Technology China Agricultural University Beijing China

Phenylalanine hydroxylase is assumed to be a key enzyme in drosopterin metabolism, but direct in vivo evidence to support this hypothesis is still *** the present study, we found a new natural reces-sive purple eye mutant of Drosophila melanogaster, Hnbp, which was a 45-nt insertion mutant in the second exon of *** insertion resulted in a predicted protein with 15 additional amino acids as compared to the wild-type *** analysis of protein structure showed that the predicted mutant protein probably had two more β-sheets, which may cause instability of two α-helices near the catalytic centre of the enzyme in the Biopterin-Hydroxyl binding *** mutant showed eye color defect with decrease of mRNA level, as well as drosopterin content *** drosopterin defect could be fully rescued by expression of wild type Henna in the Hnbp background by GMR-GAL4 UAS-Henna/UAS-Henna:Hnbp/Hnbp transgenic *** taken together, it can be concluded that the mu-tation in Henna is responsible for drosopterin reduction in mutant Hnbp, which provides key in vivo evidence to support the hypothesis that Henna is involved in drosopterin synthesis.

关键词： Drosophila melanogaster Henna gene drosopterin phenylalanine hydroxylase Hnbp

Identification of yak lactate dehydrogenase B gene variants by gene cloning

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science china(life sciences) 2008年第5期51卷 430-434页

作者： ZHENG YuCai1, zhao XingBo2, ZHOU Jing1, PIAO Ying1, JIN SuYu1, HE QingHua1, HONG Jian1, li Ning2 & wu changxin2 1 college of life science and technology, Southwest university for Nationalities, Chengdu 610041, china 2 State Key Laboratory for Agrobiotechnology and Ministry of Agriculture Key Laboratory of animal Genetics and Breeding, college of animal science and technology, china agricultural university, beijing 100094, china college of life science and technology Southwest University for Nationalities Chengdu China State Key Laboratory for Agrobiotechnology and Ministry of Agriculture Key Laboratory of animal Genetics and Breeding College of Animal Science and Technology China Agricultural University Beijing China

Native polyacrylamide gel electrophoresis showed that two types of lactate dehydrogenase (LDH) existed in yaks. Based on the electrophoresis characteristics of LDH isoenzymes, yak LDH variants were speculated to be the gene mutation on H subunit encoded by B gene. According to the mobility in electrophoresis, the fast-band LDH type was named LDH-Hf and the slow-band LDH type LDH-Hs. In order to reveal the gene alteration in yak LDH variants, total RNA was extracted from heart tissues of yaks with different LDH variants, and cDNAs of the two variants were reverse transcripted. Two variants of B genes were cloned by RT-PCR. Sequence analysis revealed that four nucleotides differed between LDH-Bf and LDH-Bs, which resulted in two amino acids alteration. By Deepview software analysis of the conformation of yak LDH1 variants and H subunit, these four nucleotides altered two amino acids that generated new hydrogen bonds to change the hydrogen bonds network, and further caused subtle conformational changes between the two LDH variants.

关键词： yak lactate dehydrogenase genetic variants gene cloning molecular modeling

Sequences and polymorphisms of exons 3 and 4 in porcine UCP2 gene

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Progress in Natural science:Materials International 2002年第7期 73-74页

作者： FANG Meiying, liU Chousheng, zhao Xingbo, li Ning and wu changxin(1. college of animal science and technology, china agricultural university, beijing 100094, china 2. National Laboratory for A-gribiotechnology, china agricultural university, beijing 100094, china 3. Gansu Institute of animal and Veterinary science, Pingliang 744000, china) college of animal science and technology，china agricultural university，beijing 100094，china National Laboratory for Agribiotechnology，china agricultural university，beijing 100094，china Gansu Institute of animal and Veterinary science，Pingliang 744000，china National Laboratory for Agribiotechnology，china agricultural university，beijing 100094，china

Uncoupling proteins are mitochondrial membrane transporters, which regulate metabolic pathways of energy balance, and are associated with biological traits of animal body weight, resting metabolic rates and energy conversion. In this study, a region of the exons 3 and 4 of pig UCP2 gene was cloned and analyzed, and a new single nucleotide polymorphic site was detected by PCR-SSCP in five pig breeds. This newfound polymorphism results from a T to G stbstitution at the position of nucleotide 272, which is located in intron3.

关键词： pig uncoupling proteins PCR-SSCP.

Effects of different states of sheep fetal fibroblasts as donor cells on the early development in vitro of reconstructed sheep embryos

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science china(life sciences) 2007年第2期50卷 178-185页

作者： WANG Hai, AO Hong, PAN QiuZhen, li RongQi, zhao MengBin, li AN ZhengXing, li Ning & wu changxin 11nstitute of Zoology, Chinese Academy of sciences, beijing 100080, china 2 Institute of animal science, Chinese Academy of agricultural sciences, beijing 100094, china 3 Northeast agricultural university, Harbin 150030, china 4beijing Glorious Land agricultural Co., Ltd. beijing 100049, china 5 college of animal science and technology, china agricultural university, beijing 100094, china 6 State Key Laboratory of Agrobiotechnology, china agricultural university, beijing 100094, china Institute of Zoology Chinese Academy of Sciences Beijing 100080 China college of animal science and technology China Agricultural University Beijing 100094 China Institute of animal science Chinese Academy of Agricultural Sciences Beijing 100094 China Northeast agricultural university Harbin 150030 China college of animal science and technology China Agricultural University Beijing 100094 China beijing Glorious Land agricultural Co. Ltd. Beijing 100049 China college of animal science and technology China Agricultural University Beijing 100094 China State Key Laboratory of Agrobiotechnology China Agricultural University Beijing 100094 China college of animal science and technology China Agricultural University Beijing 100094 China

To investigate the effects of different states of donor cells on the development of reconstructed sheep embryos, we designed five treatments of donor cells, including cell passage, cell size, serum starvation, colchicine treatment and gene transfection. Results are as follows: (Ⅰ) Compared with 16-18 passage cells, the morula/blastocyst rate of 5-7 passage cells as donor nuclei was significantly higher (17.3% vs. 4.9%, Plity to develop into morula/blastocysts than those from small or large ones. (Ⅲ) The morula/blastocyst rate of reconstructed embryos from donor cells of SS (serum starvation) was lower than that from donor cells of NSS (non-serum starvation), but no significant difference was detected between SS and NSS(11.8% vs. 18.6%, P>0.05). (Ⅳ) Fetal fibroblasts treated with 0.05μmol/L colchicine exhibited a higher morula/blastocyst rate of reconstructed embryos than those treated with 0.10 μmol/L colchicine and untreated ones (27.5% vs. 12.1%, 17.1%), however, no significant difference among the three treatments was detected (P>0.05). (Ⅴ) The morula/blastocyst rate of reconstructed embryos from fetal fibroblasts transfected with GFP gene only was 3.1%, significantly lower than that from non-transgenic cells (3.1% vs. 20.4%, Plitate the developmen

关键词： sheep nuclear transfer colchicine serum starvation fetal fibroblast GFP gene

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猪FSHβ亚基基因RFLPs研究初报

畜牧兽医学报 1998年第1期29卷 23-26页

作者：赵要风李宁陈永福吴常信中国农业大学农业生物技术国家重点实验室中国农业大学动物科技学院

本文用猪ＦＳＨβ基因ｃＤＮＡ作为探针，对二花脸猪、梅山猪、长白猪、大约克猪、香猪基因组ＤＮＡ进行ＥｃｏＲＩ、ＢａｍＨＩ和ＨｉｎｄⅢ三种限制性内切酶的Ｓｏｕｔｈｅｒｎ印迹分析。发现猪品种之间在该位点变异较大，而且发现Ｆ... 详细信息

本文用猪ＦＳＨβ基因ｃＤＮＡ作为探针，对二花脸猪、梅山猪、长白猪、大约克猪、香猪基因组ＤＮＡ进行ＥｃｏＲＩ、ＢａｍＨＩ和ＨｉｎｄⅢ三种限制性内切酶的Ｓｏｕｔｈｅｒｎ印迹分析。发现猪品种之间在该位点变异较大，而且发现ＦＳＨβ／ＢａｍＨＩＲＦＬＰｓ中，太湖猪（二花脸、梅山猪）表现出品种内一致性。

关键词：猪 FSHβ基因 RFLPs 遗传育种

The screening and identification of endogenous retrovirus free CEMPs

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science china(life sciences) 2004年第6期47卷 562-566页

作者： LU Quanzhi1,3, HAN Hongbing1,3, liAN Zhengxing1,3, li Ning2,3, ZHANG Qingcai1, zhao Zili4, PEI Dezhi1, ZHANG Xiaolan1 & wu changxin1 1. college of animal science and technology, china agricultural university, beijing 100094, china 2. State Key Laboratory of Agrobiotechnology, beijing 100094, china 3. Laboratory of animal Genetics and Breeding, Ministry of Agriculture, beijing 100094, china 4. Sangzi Hu Pig Breeding Farm, Hubei 434010, china Correspondence should be addressed to lian Zhengxing college of animal science and technology China Agricultural UniversityBeijing 100094China Laboratory of animal Genetics and Breeding Ministry of AgricultureBeijing 100094China State Key Laboratory of Agrobiotechnology Beijing 100094China Laboratory of animal Genetics and Breeding Ministry of AgricultureBeijing 100094China college of animal science and technology China Agricultural UniversityBeijing 100094China Sangzi Hu Pig Breeding Farm Hubei 434010China

The provirus DNA sequence of porcine endogenous retrovirus (PERV) distributed in the pig genome is the major obstacle that restricts the swine as the organ donors in xenotransplantation, and the copy number of PERV varies greatly among different breeds and individuals. In the experiment, 67 healthy, female Chinese Experimental Mini-Pigs (CEMPs) aged at 3—6 months were selected from the animal Husbandry Station of china agricultural university, the copy number of PERV and types of envelope protein gene (env) were then investigated by means of PCR analysis and Southern blotting. It is showed that the distribution of types of envelope protein gene in Landrace and CEMPs makes little difference, but the proportion of individuals carrying two types of envelope protein gene (env-A and env-B, which is de- noted as env-AB) is much larger than those which carry only one type of envelope protein gene (env-A or env-B). Meanwhile, two endogenous retrovirus free pigs were found for the first time during our research, and the copy number of others is relatively low, which is about 10 to 20. All the results illuminate the genetic diversity of indigenous pig breeds in china and the potential of CEMPs to serve as organ donors in xenotransplantation.

关键词： swine, CEMPs, porcine endogenous retrovirus (PERV), PCR, Southern blot.

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荷斯坦奶牛胚胎基因组遗传评估研究

畜牧兽医学报 2024年第7期55卷 2940-2950页

作者：郭子骄郑伟杰孙伟吴宝江包向男张琪贺巾锋包斯琴赵高平王子馨韩博李喜和孙东晓中国农业大学动物科学技术学院农业农村部动物遗传育种与繁殖重点实验室动物育种国家工程实验室畜禽生物育种全国重点实验室北京100193 内蒙古赛科星家畜种业与繁育生物技术研究院有限公司呼和浩特011517 国家乳业技术创新中心呼和浩特010020 内蒙古大学生命科学学院呼和浩特010021

旨在建立一种荷斯坦奶牛胚胎细胞早期基因组检测和遗传评估的方法并筛选优秀种用胚胎,通过胚胎移植技术选育优秀种牛,进一步缩短世代间隔,加快遗传进展。本研究基于4头荷斯坦种公牛克隆细胞,比较分析2细胞、5细胞的微量基因组DNA提取效... 详细信息

旨在建立一种荷斯坦奶牛胚胎细胞早期基因组检测和遗传评估的方法并筛选优秀种用胚胎,通过胚胎移植技术选育优秀种牛,进一步缩短世代间隔,加快遗传进展。本研究基于4头荷斯坦种公牛克隆细胞,比较分析2细胞、5细胞的微量基因组DNA提取效果、全基因组芯片检出率及错配率;随后提取24枚荷斯坦奶牛早期胚胎5细胞样本的基因组DNA,利用150K芯片进行基因组检测;基于大规模基因组选择参考群体,采用GBLUP方法计算每枚胚胎9个性状的基因组育种值。结果表明,2和5克隆细胞的基因组DNA浓度、纯度和相对完整性均较好;5克隆细胞样品的150K芯片检出率较高(89.54%~95.07%)、错配率较低(1.02%~4.67%)。24枚胚胎的5细胞样品基因组DNA质量、总量和相对完整性均较好,150K芯片数据检出率为92.28%~98.51%,9个性状(产奶、体型和体细胞评分)基因组评估准确性为55%~79%,平均基因组评估准确性不低于65%。综上,本研究优化并建立了奶牛胚胎少量细胞DNA提取及基因组评估技术,胚胎样本基因组检出率不低于90%,平均基因组评估准确性不低于0.65。

关键词：荷斯坦奶牛胚胎基因组选择

MALE STERIliTY 3 encodes a plant homeodomain-finger protein for male fertility in soybean

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Journal of Integrative Plant Biology 2022年第5期64卷 1076-1086页

作者： Jingjing Hou Weiwei Fan Ruirui Ma Bing li Zhihui Yuan Wenxuan Huang Yueying wu Quan Hu Chunjing lin Xingqi zhao bao Peng limei zhao Chunbao Zhang lianjun Sun State Key Laboratory of Agrobiotechnology Beijing Key Laboratory for Crop Genetic Improvementand College of Agronomy and BiotechnologyChina Agricultural UniversityBeijing 100193China Soybean Research Institute the National Engineering Research Center for SoybeanJilin Academy of Agricultural SciencesChangchun 130033China Jiamusi Branch of Heilongjiang Academy of agricultural sciences Jiamusi 154007China

Male-sterile plants are used in hybrid breeding to improve yield in soybean(Glycine max(L.)Merr.).Developing the capability to alter fertility under different environmental conditions could broaden germplasm resources and simplify hybrid ***,molecular mechanisms potentially underlying such a system in soybean were ***,using positional cloning,we identified a gene,MALE STERIliTY 3(MS3),which encodes a nuclear-localized protein containing a plant homeodomain(PHD)-finger domain.A spontaneous mutation in ms3 causing premature termination of MS3 translation and partial loss of the *** analysis indicated that MS3 knockout resulted in nonfunctional pollen and no selfpollinated pods,and RNA-seq analysis revealed that MS3 affects the expression of genes associated with carbohydrate ***,the fertility of mutant ms3 can restore under longd *** mutant could thus be used to create a new,more stable photoperiod-sensitive genic male sterility line for two-line hybrid seed production,with significant impact on hybrid breeding and production.

关键词： fertility restoration male sterililty PHD-finger photoperiod soybean