驯鹿的人工授精及分子鉴定
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摘要
我国驯鹿种群是300年前，随鄂温克人为躲避战乱而迁移到此。这一驯鹿种群在大兴安岭当地的生态环境中，经过数百年的封闭、隔离饲养，形成了其特有的生活习性，也是栖息在最南部少有的驯鹿种群。我国一直未开展有关驯鹿人工饲养的科学研究工作。目前，鄂温克猎民饲养驯鹿仍沿用多年来所形成的固有的传统模式，采用半野生饲养方式，对驯鹿进行粗放管理。把驯鹿散放在暂住点的周围，无人看管，自由采食，不进行补饲，不为其筹备饲料，不论天气多么恶劣，驯鹿仍依赖自然采食而维持其生存。驯鹿的繁殖是采用自然交配，种群处于自然淘汰状态，近交衰退严重。这使驯鹿的养殖业的发展受到了极大的限制。驯鹿人工授精技术的应用，可以极大地提高种鹿的质量，使良种基因得到最大限度地传播，种鹿利用率高；避免疾病传播；避免种公鹿争斗而造成损失。
驯鹿是唯一一种雌雄都长茸的鹿。驯鹿在传统中药中，一直被作为伪品处理。现今常用的鉴定驯鹿雌雄鹿茸方法有性状鉴定、显微鉴定、理化鉴定等，在分子水平上的鉴定一直没有人研究。驯鹿雌雄鹿茸分子鉴定的研究为茸鹿伪品鉴别试剂盒提供了进一步的完善。
我国驯鹿一直是闭锁繁育，种群退化明显。驯鹿遗传多样性的研究，为我国驯鹿的育种工作提供了分子基础，为以后的分子育种工作提供了可能，也为我们研究群体遗传和遗传分化、系统发育提供了基础资料。
在本次研究中主要分为三个部分。第一，驯鹿的人工授精。通过对11头母驯鹿人工授精的初步研究，结果同期发情和人工授精的结果都很理想，同期发情率和受精率均为100％。第二，驯鹿的分子生物学鉴定。本次试验采用分子生物学鉴定的方法，对雌、雄驯鹿茸的SRY基因进行扩增，从扩增的图像可以看到，我们采用的引物能够准确的对雌、雄驯鹿茸进行区分。应用DNAMAN4.0对驯鹿、梅花鹿、马鹿进行聚类分析，结果梅花鹿与马鹿同源性为0.99，梅花鹿、马鹿与驯鹿同源性为0.37。第三，驯鹿的遗传多样性研究。本次试验采用24个引物进行RAPD－PCR扩增，在其中筛选出14个扩增条带清晰引物。应用DNAMAN4.0分析，本实验驯鹿种群相似系数在0.584～1.000之间供试群体14条引物的相似系数平均值为0.792。
In China reindeer immigrated Daxinganling with Ewenkers together in order to avoid chaos of war three hundred years ago. Reindeer formed their special living habits under local environment after hundreds of isolated rearing, which were rare species in south. Research on artificial rearing reindeer had never been carried out in China. Now hunters in Ewenke had been rearing reindeer by semi-wild rearing way that is traditional extensive management. Reindeer searched for food freely around temporarily residential area without being watched and supplementary feed. Reindeer breed by natural mating, and which were eliminated through nature, close relative mating declined in reindeer community seriously, which limited the development of cultivating reindeer. The application of artificial insemination technique of reindeer improved the quality and availability of breeding reindeer, and disseminated fine genes most extensively, and avoided spreading disease and loss due to fighting of breeding male reindeer.
Reindeer were only one kind of deer whatever it was male or female had antler. Reindeer were considered fake thing in the traditional Chinese medicine. Now the ways of determining that it was male or female mainly included properties determine, determine while being micro-, the physics and chemistry determine and so on, molecular determine was never researched. The research of molecular determining male and female antler perfected differentiating reagent box of fake antler.
Reindeer were breed under isolation in China, reindeer community declined seriously. The research of genetic diversification of reindeer provided molecular base for breeding of reindeer in China, and provided possibility for the future molecular breeding research, and provided basis material for the research of community heredity, heredity differentiation and systematic growth.
The experiment was divided into three parts. First, artificial insemination of reindeer was carried out. Primary research results of artificial insemination of 11 reindeer show that Estrus Synchronization and artificial insemination were ideal, Estrus Synchronization and artificial insemination rate were 100%. Secondly, the molecular biology determine of reindeer was carried out. Adopting the molecular biology qualification method in the research, SRY genes of male and female antler were increased, we could see from increased picture prime adopted could differentiate female antler and male antler accurately. We use DNAMAN4.0 to structure the tree among Reindeer and Sika deer and Red deer.We can draw a conlusion that The similarity of Sika and Red deer are 0.99,that the similiraty among Reindeer,Sika deer and Red deer are 0.37.Thirdly, genetic diversification of reindeer was researched. 24 primes were increased by RAPD-PCR, from which 14 increased clear primes were sifted. I analyse the kindred of these 13 Reindeers with DNAMAN4.0,and draw a conclusion that the similiraty is between 0.584 and 1.000,that the similitude coefficient of 14 primes are 0.792.
Li Haitao
Animal Genetics and Breeding
Prof. Bai Xiujuan
驯鹿是唯一一种雌雄都长茸的鹿。驯鹿在传统中药中，一直被作为伪品处理。现今常用的鉴定驯鹿雌雄鹿茸方法有性状鉴定、显微鉴定、理化鉴定等，在分子水平上的鉴定一直没有人研究。驯鹿雌雄鹿茸分子鉴定的研究为茸鹿伪品鉴别试剂盒提供了进一步的完善。
我国驯鹿一直是闭锁繁育，种群退化明显。驯鹿遗传多样性的研究，为我国驯鹿的育种工作提供了分子基础，为以后的分子育种工作提供了可能，也为我们研究群体遗传和遗传分化、系统发育提供了基础资料。
在本次研究中主要分为三个部分。第一，驯鹿的人工授精。通过对11头母驯鹿人工授精的初步研究，结果同期发情和人工授精的结果都很理想，同期发情率和受精率均为100％。第二，驯鹿的分子生物学鉴定。本次试验采用分子生物学鉴定的方法，对雌、雄驯鹿茸的SRY基因进行扩增，从扩增的图像可以看到，我们采用的引物能够准确的对雌、雄驯鹿茸进行区分。应用DNAMAN4.0对驯鹿、梅花鹿、马鹿进行聚类分析，结果梅花鹿与马鹿同源性为0.99，梅花鹿、马鹿与驯鹿同源性为0.37。第三，驯鹿的遗传多样性研究。本次试验采用24个引物进行RAPD－PCR扩增，在其中筛选出14个扩增条带清晰引物。应用DNAMAN4.0分析，本实验驯鹿种群相似系数在0.584～1.000之间供试群体14条引物的相似系数平均值为0.792。
In China reindeer immigrated Daxinganling with Ewenkers together in order to avoid chaos of war three hundred years ago. Reindeer formed their special living habits under local environment after hundreds of isolated rearing, which were rare species in south. Research on artificial rearing reindeer had never been carried out in China. Now hunters in Ewenke had been rearing reindeer by semi-wild rearing way that is traditional extensive management. Reindeer searched for food freely around temporarily residential area without being watched and supplementary feed. Reindeer breed by natural mating, and which were eliminated through nature, close relative mating declined in reindeer community seriously, which limited the development of cultivating reindeer. The application of artificial insemination technique of reindeer improved the quality and availability of breeding reindeer, and disseminated fine genes most extensively, and avoided spreading disease and loss due to fighting of breeding male reindeer.
Reindeer were only one kind of deer whatever it was male or female had antler. Reindeer were considered fake thing in the traditional Chinese medicine. Now the ways of determining that it was male or female mainly included properties determine, determine while being micro-, the physics and chemistry determine and so on, molecular determine was never researched. The research of molecular determining male and female antler perfected differentiating reagent box of fake antler.
Reindeer were breed under isolation in China, reindeer community declined seriously. The research of genetic diversification of reindeer provided molecular base for breeding of reindeer in China, and provided possibility for the future molecular breeding research, and provided basis material for the research of community heredity, heredity differentiation and systematic growth.
The experiment was divided into three parts. First, artificial insemination of reindeer was carried out. Primary research results of artificial insemination of 11 reindeer show that Estrus Synchronization and artificial insemination were ideal, Estrus Synchronization and artificial insemination rate were 100%. Secondly, the molecular biology determine of reindeer was carried out. Adopting the molecular biology qualification method in the research, SRY genes of male and female antler were increased, we could see from increased picture prime adopted could differentiate female antler and male antler accurately. We use DNAMAN4.0 to structure the tree among Reindeer and Sika deer and Red deer.We can draw a conlusion that The similarity of Sika and Red deer are 0.99,that the similiraty among Reindeer,Sika deer and Red deer are 0.37.Thirdly, genetic diversification of reindeer was researched. 24 primes were increased by RAPD-PCR, from which 14 increased clear primes were sifted. I analyse the kindred of these 13 Reindeers with DNAMAN4.0,and draw a conclusion that the similiraty is between 0.584 and 1.000,that the similitude coefficient of 14 primes are 0.792.
Li Haitao
Animal Genetics and Breeding
Prof. Bai Xiujuan
引文
1.罗剑通.冷俊丰.宋海燕.赵蒙.郑兴涛.赵裕方.赵广华.梅花鹿人工授精实验报告.特产研究.2000.4:28-30
2.包海鹰.我国鹿类及各种鹿角形态特征的研究.中医药信息.1994.5.27-31
3.陈代贤,郭月秋,门启鸣,鹿胎及其混伪品的性壮鉴定研究,中药材,1997,20(5):226-229
4.范青,刘辉,王萍萍,郭月秋,陈代贤.聚丙烯酰胺凝胶电泳在鹿血与其他动物血鉴别中的应用.中医药学报.2000.5:39—40
5.傅文,唐双焱,陈永久,王建云,蒋序,张亚平.鹿属动物线粒体ＤＮＡ序列测定的研究.中国药学杂志 2000.12:803-804
6.龚大洁.周开亚.DNA分子系统学在爬行动物中的应用.动物学杂志.2000.4.35-53
7.郭彩玉.徐力.牛志多.鹿茸的组织学研究.特产研究.1996.3：18—20
8.郭金虎,单祥年,常青,余多慰,武景阳.赤麂ＳＲＹ基因的测序及其与部分偶蹄目动物ＳＲＹ基因序列的比较.动物学报.2001.2.:145～149
9.郭月秋,陈代贤,刘辉,巩丽云.鹿心及其伪品的免疫凝集试验鉴别探讨.中医药学报. 2000.1:62-63
10.郭月秋,陈代贤,刘辉,王萍萍,聚丙烯酰胺凝胶电泳鉴别鹿血与其它动物血,时药 .2000.3:232
11.郭月秋,陈代贤,刘辉,杨光,鹿鞭及伪品的免疫凝集试验鉴别.中国药事2000.2.:103-105
12.韩盛兰主编.鹿养殖与鹿产品加工.科学技术文献出版社.1999:16-25
13.郝春明,任国栋,鹿鞭的性状鉴别,中药材,1993,16(3):22-24
14.胡振东.郭文场主编.养鹿与鹿产品.辽宁科学技术出版社1998:27-30
15.黄璐琦,分子生药学,北京医科大学出版社,2000年,254-263
16.黄守华,李淑玲,常虹.驯鹿的饲养及繁殖. 黑龙江动物繁殖 1999(2)21-23
17.黄原著.分子系统学－原理.方法及应用 . 中国农业出版社.1998.61-65
18.李长生，王喜萍.鹿茸的加工技术.农牧产品开发2000．5：23-25
19.李和平,师守堃,李生.中国茸鹿品种 (品系 )的随机扩增多态ＤＮＡ(ＲＡＰＤ)研究.应用与环境生物学报.20 0 0 , 3:237～ 246
20.李和平.师守堃 .李生.中国茸鹿品种(品系)间遗传关系的分析 .经济动物学报 2000,4(2) :23～26
21.李明.王小明.盛和林.玉手英利.增田隆一.永田纯子.大泰司纪之.四种鹿属动物的线粒体.DNA差异和系统进化关系研究.动物学报. 1994 5( 1).99～ 105,
22.李明.王小明.盛和林.玉手英利.增田隆一.永田纯子.大泰司纪之.四种鹿属动物的线粒体DNA差异和系统进化关系的研究.动物学报.1999.1:99～105
24.刘向华,鹿类中药材的位点特异性ＰＣＲ鉴定研究, 药学学报2001,(36)8:631- 635
25.刘向华.鹿类中药材的位点特异性ＰＣＲ鉴定研究.药学学报2001,8 :631- 635
26.卢圣栋，现代分子生物学实验技术，高教出版社
27.吕碧文,俞颂东,金水仙，牛早期胚胎性别鉴定的PCR方法研究，武汉大学学报(自然科学版)，1999，4：211～214
28.任玮,邓晓健,注意驯鹿角片混充鹿茸片,中药材,1993,16(4):23-24
29.唐伯平.周开亚.宋大祥.核rDNA ITS区序列在无脊椎动物分子系统研究中的应用.动物学杂志.2002.4.:67-73
30.唐双焱,傅文,陈永久,王建云,蒋序,张亚平,中药材鹿鞭的分子鉴定研究.中国中药杂
志.2002.8:573-576
31.唐双焱.傅文.陈永久.王建云.蒋序.张亚平.鹿线粒体ＤＮＡ的种属特异性片段研究.华西药学杂志.2001.5:325～326
32.唐双焱.寻找鹿线粒体ＤＮＡ上区别于常见伪充鹿类药材来源动物的种属特异性片段.华西药学杂志.2001,5:325～ 326
33.王志军,汪世忠.鹿的品种分类及生产性能.河南畜牧兽医.1999.11:34-35
34.王宗仁. 鹿科动物血清蛋白质SDS—聚丙烯酰胺凝胶电泳分析.兽类学报.1988.8.（1）：13-20
35.何学华.马鹿人工授精技术.特种经济动植物.2001.6:5
36.熊远著主编.猪生化及分子遗传实验导论.第一版.中国农业出版社,1999:39-52
37.徐国钧.徐珞珊.中药材粉末显微鉴定.第一版.人民出版社.1986:23-29
38.徐营,杨利国,郭爱珍,杨俊杰.性别决定基因ＳＲＹ的研究进展.生物技术通报 2002.1.30-33
39.俞颂东.吕碧文,金水仙,杨继宗.应用牛的Y染色体特异引物扩增羊SRY基因的研究.浙江农业学报.1995.4：334－336
40.曾申明.朱士恩.主编.鹿的养殖疾病防治产品加工.第一版.北京中国农业出版.1999:12-22
41.张保国.李钦.鹿角.鹿茸混淆品-狍角.狍茸的鉴别研究.中国药学杂志.1992.27（12）：713—715
42.张才骏,王勇,马睿麟. 白唇鹿.马鹿和梅花鹿血清碱性磷酸酶同工酶的酶谱 . 1998.1:4-9
43.张华.马新武主编.珍稀动物性药材生产.第一版.中国农业出版社.1997:80-95
44.张敏.白秀娟.驯鹿18SrRNA基因的克隆测序.经济动物学报 .2003.1.15-17
45.赵世臻,沈广主编,中国养鹿大成,中国农业出版社,2003,第二版,13-20
46.仲昭庆.卢守奇.鹿鞭的电泳鉴别.中药材.1997.20（7）：339-340
47朱秀翔.脱氧核糖核酸分子遗传标记技术在中药鉴定中的应用.时珍国医国药.2001.10.950-953
48.魏海军.杨福合.陆纯志.杨富山.张传友.常忠娟. CIDR和PMSG诱导东北梅花鹿同期发情的49.初步研.经济动物学报.2003.7(3):27~29
50.钟立成.卢向东.李艳冰.张仁明.半散放东北马鹿人工授精技术.经济动物学报.2002.6(3):19~22
51.钟立成.卢向东.邵伟庚.张仁明. 利用人工授精技术改良半散放马鹿的研究.林业科技.2002.9:27~29
52.马泽芳.王玉龙.贺俊峰.雷振中.鹿的同期发情技术研究进展.东北林业大学学报.2000.3:71~73
53.郑兴涛.马鹿和梅花鹿人工授精技术.特种经济动植物.2002.10:8
54.武春田.张贵学.国泰.赵裕方.赵列平.马鹿假台鹿采精技术的研究.黑龙江动物繁殖.1994.2(3):22~23
55.何学华.马鹿人工授精技术.特种经济动植物.2001.6:5
56.马泽芳.田长永.刘家亮.张艳禾.崔丽春.姜晓东.崔凯.雷振中.靳占江.刘桂彬.王玉华.梅花鹿的同期发情技术.东北农业大学学报.2002.7:73~76
57.王柏林.赵裕方.张殿宇.李占武.杨立祥.马殿臣.艾永利.梅花鹿人工授精配套新技术研究.特产研究.2002.4:1~5
58.赵裕芳.茸鹿人工授精的几个关键问题及应对措施.养殖技术顾问.2003.10:38
59.赵裕芳.赵列平.茸鹿人工授精是我国鹿业发展的关键技术.养殖技术顾问.2003.9:41
60.李武.陈静波.塔里木马鹿冷冻精液制作技术的研究.草食家畜.1997.1(94):24
61.赵列平.赵裕方.武春田. 郑兴涛.邴国良.赵蒙.天山马鹿清源品系人工授精新技术及推广应用效果的研究.经济动物学报.1998.2(2):23～27
62.赵列平.赵裕方.武春田. 郑兴涛.邴国良.赵蒙.天山马鹿清原品系幼鹿培育新技术.经济动物学报. 1998.2(4):46～52
63.赵列平.王全凯.李彦江.王忠武.刘文辉.王景武.同期发情技术在梅花鹿繁殖上的应用.经济动物学报.2003.7(3):24～26
64.张一玲.渊锡藩.尹海科.王忠林.魏志杰.山羊同期发情技术的应用.畜牧兽医杂志.2000.5(19):21~24
65.庞训胜.王荣祥.吴义.常春芳.李业厂.邓英状.孕酮阴道栓对山羊同期发情的影响.安徽技术师范学院学报.2002.16(1):7～9
66. 史宪伟,陈永久,刘爱华,等.云南4个马品种的随机扩增多态ＤＮＡ(ＲＡＰＤ)分析[Ｊ].畜牧兽医学报,1998,29(3):193-203.
67. 陈运棣,杨关福,张细权,等.用ＲＡＰＤ标记分析枫叶鸭群体的遗传结构[Ａ].中国畜禽遗传标记研究会.第六届畜禽遗传标记研讨会论文集[Ｃ].北京:中国农业出版社,1998.103-106.(004)
68.Emerson.bc.eyal.geneticanalysis of erolutionary relationships among deer(subfamily cervinae)Jheredity,1993,84(4):266~273
69.Flerov KK.Moyphology and ecology of cervidae during its evolution .J .Paleontology Translation.1957.1-2：2-16
70.Geist V,1991 Taxonomy:on an objective definition of subspecies,taxa as legal entities,and its application to Ramgifer tarandus Lin,1785 In:Butler CE, Mahoney SP,eds Proc Fourth North American Caribou Works hio St John’s Newf 1-36.
71.Geist V,1992 Endangered species and the law Nature,357:274-276.
72.Gubbay J,et al.Nature , 1990, 346:245～ 250.
73.HagelbergE,AncientboneDNAamplified,Nature,1989,342:485.
74.Hao C M ，Ren GD，Trait identification of deer penis and testis，J Chin Med Mater，1993，16，（3）：22-24
75.Hashimoto A,Studieson“singal”stiuen for evaluation of animal crude drugs.IV:Application of DNA analytical technique to genuine evalution of medicines containing animal crude drugs,Nat Med ,1998,52(1):38-46
76.Jacobs PA,et al.Nature,1966, 210:352～ 355.
59.JiaYJ，Zhou LS，Trait identification of deer tendon and its adulterants，J Chin Med Mater，1994，17（7）：17-18
77.Li M sheng HL,Tamate H, Masuda R,Nagata J(Ohtaishi N.MtDNA defference and molecular phylogeny among muskdeer,Chinese waterdeer,muntjak and deer.Acta Theriologica Sinica.1998, 18(3):184~91.
78.Li,Wang XM ,ShengHL,et al.1998,Origin and genetic diversity of four subspecies of red deer (Cervus elaphus),Zool,res,19(3):177-183.
79.LiHP,Shi SK ,Li S ,2000 ,Genetic diversity detected by RAPD in five velvet-deer breeds or strains of china ,China J Appl,Environ,Biol,6(3):237-246.
80.Lovell-badge R,et al .phil.trans R,soc.Lond,1995, 350( 1333):205～ 214.
81.Ohtaishi N,Gao Y.T. The distribution of alls pecies of deer(Tragulidae,Moschidae Cervidae) in China.Mammal Review. 1990,201:125～144.
82.Ohtaishi N.1992 The origins and evolution of the deer in China (A).In:Sfeg H L ,et
al,The Deer in China Shang Hai :East Chaina Normal University press,8-16.
83.Rusk，S.A,Spiegel,F.W,and Lee,S.B.1995.Design of polymerase chain .reaction primers for amplifying nuclear.ribosomal DNA from slima molds Mycologid 87(1)140-143
84.Sheng HL ,et al,1992,The deer china ,shanghai,East China Normal University Press
85.Sinclair AH,et al.Nature,1990,346:240～ 244.
86.Wang JY，He GX，Fu W，et al，Extraction and sequencing of trace DNA from Cornu Cervi pantotrichum，Chin Pharm J，1997，22（10）：579-583
87.Wang,Y,1996.Comparison of method of direct cloning of PCR product.Prog.Bioengineering. 16(5):55~57.
88.WangZR,Du RF,1982,Evolution of karyotype of the genus Cervus ,Acta Genetica Sinica ,9,(11):24-31.
89.White,T.J,Bruns,T,Lee,S and Taylor,J.1990.Amplification and direct sequencing of fungal ribosomal RNA genes for Phylogeneties PCR Protocools:A Guide to methods and application.(ed by M A.Innis,D.H.Gelfand,J.J Snisky,T.J.White)PP.315-322 Academil Press,San diego, California
90.Xu X ,The complete mitochondrial DNA sequence of the horse ,Equus caballus:extensitive hetropalsmy of the control region[J].Gene,1994,148(2):357.
91.XU Yan-chun, Status of microsatellites as genetic markers in cervids , Journal of Forestry Research, 2001,1: 55-58
92.Xu,J.L.and M.Xu,1998,Sex determination and sex reversal of mammal Prog.Biochem.Biophys.25(1):30~35.
93.Yang XG.Study on PCR identification of the Chinese drugs Oviductus rana and the molecular systematics of Rana in China [D] Master Dissertation[C] (in Chinese ),Nanjing Normal University,2000.22~33.
94.Zhang BG，LiQ，Liu D。Identification and study of the pseudo articles of Cornu Cervi and Cornu Cervi pantotrichum---Cornu Caperoli and Cornu Capreoli Pantotrichum，Chin Pharm J，1992，27（12）：713-715
95.Zhang,S.Z.and R.J.Zhou,1994,PCR and cloning of the Giant Panda SRY gene.Actagenetica Sinica.21(4):281～286.
96.Zhou,R.J.,Y.Q.Guo and H.H.cheng,1997,The phylogenetic comparative tree of SRY and SOX gene.Acta Zool.Sin. 43(2):192～196.
97. Capel B ,et al.Cell, 1993, 73(5) :1019～1030
98.Cronin AM,1993 Mitochondrial DNA in wildlife taxonomy and conservation biology:Cautionary notes Wild L Soc Bull, 21:339-343.
2.包海鹰.我国鹿类及各种鹿角形态特征的研究.中医药信息.1994.5.27-31
3.陈代贤,郭月秋,门启鸣,鹿胎及其混伪品的性壮鉴定研究,中药材,1997,20(5):226-229
4.范青,刘辉,王萍萍,郭月秋,陈代贤.聚丙烯酰胺凝胶电泳在鹿血与其他动物血鉴别中的应用.中医药学报.2000.5:39—40
5.傅文,唐双焱,陈永久,王建云,蒋序,张亚平.鹿属动物线粒体ＤＮＡ序列测定的研究.中国药学杂志 2000.12:803-804
6.龚大洁.周开亚.DNA分子系统学在爬行动物中的应用.动物学杂志.2000.4.35-53
7.郭彩玉.徐力.牛志多.鹿茸的组织学研究.特产研究.1996.3：18—20
8.郭金虎,单祥年,常青,余多慰,武景阳.赤麂ＳＲＹ基因的测序及其与部分偶蹄目动物ＳＲＹ基因序列的比较.动物学报.2001.2.:145～149
9.郭月秋,陈代贤,刘辉,巩丽云.鹿心及其伪品的免疫凝集试验鉴别探讨.中医药学报. 2000.1:62-63
10.郭月秋,陈代贤,刘辉,王萍萍,聚丙烯酰胺凝胶电泳鉴别鹿血与其它动物血,时药 .2000.3:232
11.郭月秋,陈代贤,刘辉,杨光,鹿鞭及伪品的免疫凝集试验鉴别.中国药事2000.2.:103-105
12.韩盛兰主编.鹿养殖与鹿产品加工.科学技术文献出版社.1999:16-25
13.郝春明,任国栋,鹿鞭的性状鉴别,中药材,1993,16(3):22-24
14.胡振东.郭文场主编.养鹿与鹿产品.辽宁科学技术出版社1998:27-30
15.黄璐琦,分子生药学,北京医科大学出版社,2000年,254-263
16.黄守华,李淑玲,常虹.驯鹿的饲养及繁殖. 黑龙江动物繁殖 1999(2)21-23
17.黄原著.分子系统学－原理.方法及应用 . 中国农业出版社.1998.61-65
18.李长生，王喜萍.鹿茸的加工技术.农牧产品开发2000．5：23-25
19.李和平,师守堃,李生.中国茸鹿品种 (品系 )的随机扩增多态ＤＮＡ(ＲＡＰＤ)研究.应用与环境生物学报.20 0 0 , 3:237～ 246
20.李和平.师守堃 .李生.中国茸鹿品种(品系)间遗传关系的分析 .经济动物学报 2000,4(2) :23～26
21.李明.王小明.盛和林.玉手英利.增田隆一.永田纯子.大泰司纪之.四种鹿属动物的线粒体.DNA差异和系统进化关系研究.动物学报. 1994 5( 1).99～ 105,
22.李明.王小明.盛和林.玉手英利.增田隆一.永田纯子.大泰司纪之.四种鹿属动物的线粒体DNA差异和系统进化关系的研究.动物学报.1999.1:99～105
24.刘向华,鹿类中药材的位点特异性ＰＣＲ鉴定研究, 药学学报2001,(36)8:631- 635
25.刘向华.鹿类中药材的位点特异性ＰＣＲ鉴定研究.药学学报2001,8 :631- 635
26.卢圣栋，现代分子生物学实验技术，高教出版社
27.吕碧文,俞颂东,金水仙，牛早期胚胎性别鉴定的PCR方法研究，武汉大学学报(自然科学版)，1999，4：211～214
28.任玮,邓晓健,注意驯鹿角片混充鹿茸片,中药材,1993,16(4):23-24
29.唐伯平.周开亚.宋大祥.核rDNA ITS区序列在无脊椎动物分子系统研究中的应用.动物学杂志.2002.4.:67-73
30.唐双焱,傅文,陈永久,王建云,蒋序,张亚平,中药材鹿鞭的分子鉴定研究.中国中药杂
志.2002.8:573-576
31.唐双焱.傅文.陈永久.王建云.蒋序.张亚平.鹿线粒体ＤＮＡ的种属特异性片段研究.华西药学杂志.2001.5:325～326
32.唐双焱.寻找鹿线粒体ＤＮＡ上区别于常见伪充鹿类药材来源动物的种属特异性片段.华西药学杂志.2001,5:325～ 326
33.王志军,汪世忠.鹿的品种分类及生产性能.河南畜牧兽医.1999.11:34-35
34.王宗仁. 鹿科动物血清蛋白质SDS—聚丙烯酰胺凝胶电泳分析.兽类学报.1988.8.（1）：13-20
35.何学华.马鹿人工授精技术.特种经济动植物.2001.6:5
36.熊远著主编.猪生化及分子遗传实验导论.第一版.中国农业出版社,1999:39-52
37.徐国钧.徐珞珊.中药材粉末显微鉴定.第一版.人民出版社.1986:23-29
38.徐营,杨利国,郭爱珍,杨俊杰.性别决定基因ＳＲＹ的研究进展.生物技术通报 2002.1.30-33
39.俞颂东.吕碧文,金水仙,杨继宗.应用牛的Y染色体特异引物扩增羊SRY基因的研究.浙江农业学报.1995.4：334－336
40.曾申明.朱士恩.主编.鹿的养殖疾病防治产品加工.第一版.北京中国农业出版.1999:12-22
41.张保国.李钦.鹿角.鹿茸混淆品-狍角.狍茸的鉴别研究.中国药学杂志.1992.27（12）：713—715
42.张才骏,王勇,马睿麟. 白唇鹿.马鹿和梅花鹿血清碱性磷酸酶同工酶的酶谱 . 1998.1:4-9
43.张华.马新武主编.珍稀动物性药材生产.第一版.中国农业出版社.1997:80-95
44.张敏.白秀娟.驯鹿18SrRNA基因的克隆测序.经济动物学报 .2003.1.15-17
45.赵世臻,沈广主编,中国养鹿大成,中国农业出版社,2003,第二版,13-20
46.仲昭庆.卢守奇.鹿鞭的电泳鉴别.中药材.1997.20（7）：339-340
47朱秀翔.脱氧核糖核酸分子遗传标记技术在中药鉴定中的应用.时珍国医国药.2001.10.950-953
48.魏海军.杨福合.陆纯志.杨富山.张传友.常忠娟. CIDR和PMSG诱导东北梅花鹿同期发情的49.初步研.经济动物学报.2003.7(3):27~29
50.钟立成.卢向东.李艳冰.张仁明.半散放东北马鹿人工授精技术.经济动物学报.2002.6(3):19~22
51.钟立成.卢向东.邵伟庚.张仁明. 利用人工授精技术改良半散放马鹿的研究.林业科技.2002.9:27~29
52.马泽芳.王玉龙.贺俊峰.雷振中.鹿的同期发情技术研究进展.东北林业大学学报.2000.3:71~73
53.郑兴涛.马鹿和梅花鹿人工授精技术.特种经济动植物.2002.10:8
54.武春田.张贵学.国泰.赵裕方.赵列平.马鹿假台鹿采精技术的研究.黑龙江动物繁殖.1994.2(3):22~23
55.何学华.马鹿人工授精技术.特种经济动植物.2001.6:5
56.马泽芳.田长永.刘家亮.张艳禾.崔丽春.姜晓东.崔凯.雷振中.靳占江.刘桂彬.王玉华.梅花鹿的同期发情技术.东北农业大学学报.2002.7:73~76
57.王柏林.赵裕方.张殿宇.李占武.杨立祥.马殿臣.艾永利.梅花鹿人工授精配套新技术研究.特产研究.2002.4:1~5
58.赵裕芳.茸鹿人工授精的几个关键问题及应对措施.养殖技术顾问.2003.10:38
59.赵裕芳.赵列平.茸鹿人工授精是我国鹿业发展的关键技术.养殖技术顾问.2003.9:41
60.李武.陈静波.塔里木马鹿冷冻精液制作技术的研究.草食家畜.1997.1(94):24
61.赵列平.赵裕方.武春田. 郑兴涛.邴国良.赵蒙.天山马鹿清源品系人工授精新技术及推广应用效果的研究.经济动物学报.1998.2(2):23～27
62.赵列平.赵裕方.武春田. 郑兴涛.邴国良.赵蒙.天山马鹿清原品系幼鹿培育新技术.经济动物学报. 1998.2(4):46～52
63.赵列平.王全凯.李彦江.王忠武.刘文辉.王景武.同期发情技术在梅花鹿繁殖上的应用.经济动物学报.2003.7(3):24～26
64.张一玲.渊锡藩.尹海科.王忠林.魏志杰.山羊同期发情技术的应用.畜牧兽医杂志.2000.5(19):21~24
65.庞训胜.王荣祥.吴义.常春芳.李业厂.邓英状.孕酮阴道栓对山羊同期发情的影响.安徽技术师范学院学报.2002.16(1):7～9
66. 史宪伟,陈永久,刘爱华,等.云南4个马品种的随机扩增多态ＤＮＡ(ＲＡＰＤ)分析[Ｊ].畜牧兽医学报,1998,29(3):193-203.
67. 陈运棣,杨关福,张细权,等.用ＲＡＰＤ标记分析枫叶鸭群体的遗传结构[Ａ].中国畜禽遗传标记研究会.第六届畜禽遗传标记研讨会论文集[Ｃ].北京:中国农业出版社,1998.103-106.(004)
68.Emerson.bc.eyal.geneticanalysis of erolutionary relationships among deer(subfamily cervinae)Jheredity,1993,84(4):266~273
69.Flerov KK.Moyphology and ecology of cervidae during its evolution .J .Paleontology Translation.1957.1-2：2-16
70.Geist V,1991 Taxonomy:on an objective definition of subspecies,taxa as legal entities,and its application to Ramgifer tarandus Lin,1785 In:Butler CE, Mahoney SP,eds Proc Fourth North American Caribou Works hio St John’s Newf 1-36.
71.Geist V,1992 Endangered species and the law Nature,357:274-276.
72.Gubbay J,et al.Nature , 1990, 346:245～ 250.
73.HagelbergE,AncientboneDNAamplified,Nature,1989,342:485.
74.Hao C M ，Ren GD，Trait identification of deer penis and testis，J Chin Med Mater，1993，16，（3）：22-24
75.Hashimoto A,Studieson“singal”stiuen for evaluation of animal crude drugs.IV:Application of DNA analytical technique to genuine evalution of medicines containing animal crude drugs,Nat Med ,1998,52(1):38-46
76.Jacobs PA,et al.Nature,1966, 210:352～ 355.
59.JiaYJ，Zhou LS，Trait identification of deer tendon and its adulterants，J Chin Med Mater，1994，17（7）：17-18
77.Li M sheng HL,Tamate H, Masuda R,Nagata J(Ohtaishi N.MtDNA defference and molecular phylogeny among muskdeer,Chinese waterdeer,muntjak and deer.Acta Theriologica Sinica.1998, 18(3):184~91.
78.Li,Wang XM ,ShengHL,et al.1998,Origin and genetic diversity of four subspecies of red deer (Cervus elaphus),Zool,res,19(3):177-183.
79.LiHP,Shi SK ,Li S ,2000 ,Genetic diversity detected by RAPD in five velvet-deer breeds or strains of china ,China J Appl,Environ,Biol,6(3):237-246.
80.Lovell-badge R,et al .phil.trans R,soc.Lond,1995, 350( 1333):205～ 214.
81.Ohtaishi N,Gao Y.T. The distribution of alls pecies of deer(Tragulidae,Moschidae Cervidae) in China.Mammal Review. 1990,201:125～144.
82.Ohtaishi N.1992 The origins and evolution of the deer in China (A).In:Sfeg H L ,et
al,The Deer in China Shang Hai :East Chaina Normal University press,8-16.
83.Rusk，S.A,Spiegel,F.W,and Lee,S.B.1995.Design of polymerase chain .reaction primers for amplifying nuclear.ribosomal DNA from slima molds Mycologid 87(1)140-143
84.Sheng HL ,et al,1992,The deer china ,shanghai,East China Normal University Press
85.Sinclair AH,et al.Nature,1990,346:240～ 244.
86.Wang JY，He GX，Fu W，et al，Extraction and sequencing of trace DNA from Cornu Cervi pantotrichum，Chin Pharm J，1997，22（10）：579-583
87.Wang,Y,1996.Comparison of method of direct cloning of PCR product.Prog.Bioengineering. 16(5):55~57.
88.WangZR,Du RF,1982,Evolution of karyotype of the genus Cervus ,Acta Genetica Sinica ,9,(11):24-31.
89.White,T.J,Bruns,T,Lee,S and Taylor,J.1990.Amplification and direct sequencing of fungal ribosomal RNA genes for Phylogeneties PCR Protocools:A Guide to methods and application.(ed by M A.Innis,D.H.Gelfand,J.J Snisky,T.J.White)PP.315-322 Academil Press,San diego, California
90.Xu X ,The complete mitochondrial DNA sequence of the horse ,Equus caballus:extensitive hetropalsmy of the control region[J].Gene,1994,148(2):357.
91.XU Yan-chun, Status of microsatellites as genetic markers in cervids , Journal of Forestry Research, 2001,1: 55-58
92.Xu,J.L.and M.Xu,1998,Sex determination and sex reversal of mammal Prog.Biochem.Biophys.25(1):30~35.
93.Yang XG.Study on PCR identification of the Chinese drugs Oviductus rana and the molecular systematics of Rana in China [D] Master Dissertation[C] (in Chinese ),Nanjing Normal University,2000.22~33.
94.Zhang BG，LiQ，Liu D。Identification and study of the pseudo articles of Cornu Cervi and Cornu Cervi pantotrichum---Cornu Caperoli and Cornu Capreoli Pantotrichum，Chin Pharm J，1992，27（12）：713-715
95.Zhang,S.Z.and R.J.Zhou,1994,PCR and cloning of the Giant Panda SRY gene.Actagenetica Sinica.21(4):281～286.
96.Zhou,R.J.,Y.Q.Guo and H.H.cheng,1997,The phylogenetic comparative tree of SRY and SOX gene.Acta Zool.Sin. 43(2):192～196.
97. Capel B ,et al.Cell, 1993, 73(5) :1019～1030
98.Cronin AM,1993 Mitochondrial DNA in wildlife taxonomy and conservation biology:Cautionary notes Wild L Soc Bull, 21:339-343.
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