13/17染色体易位纯合子猪群随机扩增多态性DNA指纹分析
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摘要
本研究利用随机扩增多态性DNA（RAPD）技术，对13/17染色体易位
纯合子猪（2n=36）、13/17染色体易位条合子猪（2n=37）及正常核型猪
（2n=38）三个群体及14个个体的遗传结构进行了分析。在所采用的60
条随机引物中有30条可获得清晰的扩增产物，其中22条引物产生多态性
片段，而且其多态性较丰富，其余8条均产生的是单态性片段。不同引物
所扩增出的条带数目、片段大小有明显不同，每条引物扩增出的条带数介
于6～14之间，平均为11.4条，片段大小为200～4000bP。根据RAPD的
结果计算出三个群体及14个个体间的共有带率F及遗传距离指数D，三个
群体的共有带率F介于0.988～0.967之间，遗传距离指数D介于0.012～
0.033之间，其中易位纯合子猪群与正常核型猪群之间遗传距离最大，为
0.033，易位杂合子猪群与正常核型猪群之间遗传距离为0.028，易位纯合
子猪群与易位杂合子猪群之间遗传距离最小，为0.012，14个个体间的共
有带率F介于0.961～0.990之间，遗传距离指数D介于0.010～0.039之
间。在此基础上绘制出三个群体及14个个体的亲缘关系树状聚类图。遗
传距离指数的计算结果及由此所绘制的亲缘关系树状聚类图均表明了群
体间及个体间亲缘关系的远近程度，统计结果与实际情况相符。由RAPD
的结果统计出三个群体的遗传多样性指数Ho。Ho在三个群体中的分布有
明显差异。其中在易位纯合子猪群内最小（0～0.174，平均为0.084），在
正常核型猪群内最大（0.078～0.198，平均为0.138），在易位杂合子猪群
内（0～0.176，平均为0.109）介于二者之间。遗传距离指数及遗传多样
性指数对群体遗传变异程度的统计结果一致。通过群体内和群体间变异所
占比例的计算结果可见：不同引物所引起的群体内变异所占比例普遍小于
群体间变异所占比例，表明该猪群的变异主要来源于群体间的变异，群体
间的变异大于群体内的变异。t检验的结果表明，正常核型猪群体与易位
杂合子猪、易位纯合子猪群体之间的变异程度有显著差异（p＜0.05），而
易位纯合子猪与易位杂合子猪群体之间的变异程度无显著差异（p＞0.05）。
＋文扫昙W17 W色体易位此旮予精瘁闪机扩增乡足泣皿A指政艾折
一
在本次实验中发现，由引物0PQ14扩增出一条265hp的特异性片段，此片
段只出现于正常核型猪群体及个体的扩增结果内，而未出现于易位纯合于
猪与易位杂合子猪群体及个体的扩增结果内。该片段很可能为13／17染色
体易位过程中丢失的片段。将此片段进行了口收、克隆及序列测定，为进
一步的分于遗传学研究奠定了基础。同时本研究得到了以下结论：在相同
条件下利用相同引物由混合DNA池作为模板而获得的扩增产物的结果较个
体DNA模板效果好；进行群体间遗传关系分析时，群体内取样大小对统计
分析结果影响不大，其结果主要耿决于引物数目的多少：叫m结果的可信
度取决于多方面因素，包含反应体系内各物质的组成情况、扩增反应条件、
电泳检测条件及一些主观因素的影响。
The homozygous pig population for 13/17 Robertsonian translocation
with 2n=36, XY or XX rob (13;17) is a new genetic type in the domestic pig.
which was obtained by means of mating the male and female heterozygous pig
of 13/17 Robertsonian translocation. Random amplified polymorphic DNA
(RAPD) techniques were used to investigate the genetic structures of the
homozygous 13/17 Robertsonian translocation pig population, the
heterozygous 13/17 Robertsonian translocation pig population and the normal
pig population. Three DNA pools of the populations and 14 individual DNA
samples were used. Sixty random primers were screened, of which, amplified
result from 22 primers were polymorphism while the other 8 were
monomorphism. The fragment length of products from 30 primers in different
samples were between 200 and 4000 bp and the band numbers were between 6
and 14, which were distinctly and easy to be distinguished. The fragment
shared (F) and the genetic distance (D) was calculated based on it. The genetic
distance indices matrixes indicated that it was between 0.012 and 0.033 in 3
populations and between 0.010 and 0.039 in 14 individuals, the genetic
distance indices between and within the population of the 13/17 Robertsonian
translocation were low, The polygenetic trees ~vere drawn, which showed the
genetic relationship among 3 populations and 14 individuals objectively and
explicitly. The genetic diversity index (Ho) illustrated as that: Ho in the normal
pig population was high (0.078~-0.198). while it was low in the heterozygous
for 13/17 Robertsonian translocation pig population (0-.-0.176) and lower in the
homozygous for 13/17 Robertsonian translocation pig population (0~0.174).
The genetic variation among the populations is greater than that within the
populations. The genetic diversity index (Ho) of the homozygous and
heterozygous for 13/17 Robertsonian pig translocations is significantly lower
than that of the normal pig population (p<0.0 5), while the difference between
the homozygous pig population and the heterozygous pig population is not
significant (p>0.O5). It was concluded that the similarity conclusion on
heredity and variation in populations by the genetic distance indices and the
genetic diversity indices, and it was convenient and. feasible and effective in
47
13/17 ~4*~#~ IL*~*i~.~. DNA j~~~4Jj
closely genetic related. In the analysis b~?RAPD, a normal pigs specific
fragment (265bp) was found, which could not he amplified in the other
Robertsonia translocation populations; it indicated that it might be the lost
fragment during the formation of 13/17 Robcrtsonian translocation. The
specific amplified fragment has been cloned and sequenced with pMDI 8桾
vector, it would be the foundation of further molecular genetic marker
research.
纯合子猪（2n=36）、13/17染色体易位条合子猪（2n=37）及正常核型猪
（2n=38）三个群体及14个个体的遗传结构进行了分析。在所采用的60
条随机引物中有30条可获得清晰的扩增产物，其中22条引物产生多态性
片段，而且其多态性较丰富，其余8条均产生的是单态性片段。不同引物
所扩增出的条带数目、片段大小有明显不同，每条引物扩增出的条带数介
于6～14之间，平均为11.4条，片段大小为200～4000bP。根据RAPD的
结果计算出三个群体及14个个体间的共有带率F及遗传距离指数D，三个
群体的共有带率F介于0.988～0.967之间，遗传距离指数D介于0.012～
0.033之间，其中易位纯合子猪群与正常核型猪群之间遗传距离最大，为
0.033，易位杂合子猪群与正常核型猪群之间遗传距离为0.028，易位纯合
子猪群与易位杂合子猪群之间遗传距离最小，为0.012，14个个体间的共
有带率F介于0.961～0.990之间，遗传距离指数D介于0.010～0.039之
间。在此基础上绘制出三个群体及14个个体的亲缘关系树状聚类图。遗
传距离指数的计算结果及由此所绘制的亲缘关系树状聚类图均表明了群
体间及个体间亲缘关系的远近程度，统计结果与实际情况相符。由RAPD
的结果统计出三个群体的遗传多样性指数Ho。Ho在三个群体中的分布有
明显差异。其中在易位纯合子猪群内最小（0～0.174，平均为0.084），在
正常核型猪群内最大（0.078～0.198，平均为0.138），在易位杂合子猪群
内（0～0.176，平均为0.109）介于二者之间。遗传距离指数及遗传多样
性指数对群体遗传变异程度的统计结果一致。通过群体内和群体间变异所
占比例的计算结果可见：不同引物所引起的群体内变异所占比例普遍小于
群体间变异所占比例，表明该猪群的变异主要来源于群体间的变异，群体
间的变异大于群体内的变异。t检验的结果表明，正常核型猪群体与易位
杂合子猪、易位纯合子猪群体之间的变异程度有显著差异（p＜0.05），而
易位纯合子猪与易位杂合子猪群体之间的变异程度无显著差异（p＞0.05）。
＋文扫昙W17 W色体易位此旮予精瘁闪机扩增乡足泣皿A指政艾折
一
在本次实验中发现，由引物0PQ14扩增出一条265hp的特异性片段，此片
段只出现于正常核型猪群体及个体的扩增结果内，而未出现于易位纯合于
猪与易位杂合子猪群体及个体的扩增结果内。该片段很可能为13／17染色
体易位过程中丢失的片段。将此片段进行了口收、克隆及序列测定，为进
一步的分于遗传学研究奠定了基础。同时本研究得到了以下结论：在相同
条件下利用相同引物由混合DNA池作为模板而获得的扩增产物的结果较个
体DNA模板效果好；进行群体间遗传关系分析时，群体内取样大小对统计
分析结果影响不大，其结果主要耿决于引物数目的多少：叫m结果的可信
度取决于多方面因素，包含反应体系内各物质的组成情况、扩增反应条件、
电泳检测条件及一些主观因素的影响。
The homozygous pig population for 13/17 Robertsonian translocation
with 2n=36, XY or XX rob (13;17) is a new genetic type in the domestic pig.
which was obtained by means of mating the male and female heterozygous pig
of 13/17 Robertsonian translocation. Random amplified polymorphic DNA
(RAPD) techniques were used to investigate the genetic structures of the
homozygous 13/17 Robertsonian translocation pig population, the
heterozygous 13/17 Robertsonian translocation pig population and the normal
pig population. Three DNA pools of the populations and 14 individual DNA
samples were used. Sixty random primers were screened, of which, amplified
result from 22 primers were polymorphism while the other 8 were
monomorphism. The fragment length of products from 30 primers in different
samples were between 200 and 4000 bp and the band numbers were between 6
and 14, which were distinctly and easy to be distinguished. The fragment
shared (F) and the genetic distance (D) was calculated based on it. The genetic
distance indices matrixes indicated that it was between 0.012 and 0.033 in 3
populations and between 0.010 and 0.039 in 14 individuals, the genetic
distance indices between and within the population of the 13/17 Robertsonian
translocation were low, The polygenetic trees ~vere drawn, which showed the
genetic relationship among 3 populations and 14 individuals objectively and
explicitly. The genetic diversity index (Ho) illustrated as that: Ho in the normal
pig population was high (0.078~-0.198). while it was low in the heterozygous
for 13/17 Robertsonian translocation pig population (0-.-0.176) and lower in the
homozygous for 13/17 Robertsonian translocation pig population (0~0.174).
The genetic variation among the populations is greater than that within the
populations. The genetic diversity index (Ho) of the homozygous and
heterozygous for 13/17 Robertsonian pig translocations is significantly lower
than that of the normal pig population (p<0.0 5), while the difference between
the homozygous pig population and the heterozygous pig population is not
significant (p>0.O5). It was concluded that the similarity conclusion on
heredity and variation in populations by the genetic distance indices and the
genetic diversity indices, and it was convenient and. feasible and effective in
47
13/17 ~4*~#~ IL*~*i~.~. DNA j~~~4Jj
closely genetic related. In the analysis b~?RAPD, a normal pigs specific
fragment (265bp) was found, which could not he amplified in the other
Robertsonia translocation populations; it indicated that it might be the lost
fragment during the formation of 13/17 Robcrtsonian translocation. The
specific amplified fragment has been cloned and sequenced with pMDI 8桾
vector, it would be the foundation of further molecular genetic marker
research.
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