Materials & Methods1Department and Graduate Institute of
Forensic Medicine, College of Medicine, National
Taiwan University 2Department of Obstetrics and Gynecology,
National Taiwan University Hospital and
College of Medicine, National Taiwan University 3Department of
Medical Genetics, National Taiwan University Hospital
4Genephile Bioscience Laboratory, Taipei, Taiwan
Correspondence to: Professor James Chun-I Lee Telephone No:
+886-2-2312-3456 ext. 5504
Fax No: +886-2-8369-3450 E-mail address:
[email protected]
Abstract
The cytochrome B gene (MTCYB) within the mitochondrial genome has
been widely used in species identification and taxonomic research.
In this study, the population variation of the MTCYB gene in humans
was demonstrated. The sequence polymorphism of the MTCYB gene was
determined in 417 subjects in 8 populations in Taiwan (Taiwan
Chinese, mainland Chinese, Filipino, Thai, Vietnamese, and
Caucasian). Single nucleotide polymorphisms of the populations and
the genetic distance between these populations were analyzed. There
were 108 variable positions with a total of 99 haplotypes. The
distribution of varied sequences from the revised Cambridge
Reference Sequence (rCRS) was from 1 to 9 positions, with an
average of 4.26. Discrimination powers of the MTCYB gene in the 8
populations, ranging from 0.7888 (Tao) to 0.9224 (Filipino), were
observed. A population-specific position of the MTCYB gene was
noted in Tao and Caucasian populations in this study. There were
statistically significant differences of genetic distance between
Taiwan Chinese and Caucasian, between Taiwan Chinese and Tao, and
between Taiwan Chinese and Filipino. A phylogenic tree using the
neighbor-joining method based on the MTCYB gene sequence of these
populations was constructed, which grouped the Taiwan Chinese,
indigenous Taiwanese, mainland Chinese, Thais, and Vietnamese in
the same cluster. In conclusion, there are sufficient sequence
polymorphisms of the MTCYB gene in individuals of different
populations, which can be used in the identification of human
ethnic groups, and the analyses of forensic casework.
Key Words: mitochondrial DNA, MTCYB gene, sequence
polymorphism
1
87 96
14,733 10,636 4,081 16
87 7.4% 96 10.8%
40%
166
2.4-3.0
25% 30%
50%(17.3%)
10% 35-44
45-54 93
A-2
mailto:
[email protected]
2.70 9.15 /
36.3551.72 20.58
2.553.10 6.44 /
1
DNA DNA
(National Institute of Health,
USA) NCBI (National Center for Biotechnology Information) DNA
B (Cytochrome B) DNA
8 DNA PCR
DNA
DNA PCR 111bp-
158bp-228bp-273bp-334bp-397bp-438bp- 468bp-
DNA PCR DNA NCBI DNA
DNA 8
DNA-PCR
A-4
E-mail*:
[email protected] Tel: +61 432067779
Abstract
Palynology, the study of pollen, spores and other acid-resistant
microscopic plant bodies
collectively known as palynomorphs, has been used as a geological,
botanical, geographical and
archaeological tool since the early 20th century, but has only
recently emerged as a forensic tool.
Pollen and spores are produced in vast numbers and are abundant in
soil and dust, they are
microscopic in size, resistant to decay, and can be morphologically
identified to the plant species that
produced them. Palynomorph assemblages that occur in soil from any
one locality can be
distinguished from assemblages found in soil only metres away, and
from those found in soil from
other places and even different countries.
Palynomorph assemblages from soil and water at a crime scene or
area of interest can be compared
with those found in dust, dirt or soil from people or items of
interest to prove or disprove a relationship
between them. In addition, they can also help determine the
geographic origin of items of interest
including illicit drugs and illegal imports. Although palynology
can be used to help bring about a
conviction, it can be especially useful in suggesting what
direction an investigation should take, and as
corroborative evidence.
Forensic palynology is commonly used in New Zealand and the United
Kingdom and now
becoming used more often in Australia and the United States of
America, but elsewhere in the world
its application is more novel than routine. Why? Because those who
could use it know little or nothing
about it. It is time to widely educate those who could use it. We
will briefly outline how palynology
can be used as a forensic tool, illustrated with case histories
including how palynology has helped to
catch a murderer, disprove a rapist’s alibi and identify a robber,
and how cannabis can leave evidence
on clothing.
28 11 20
16 7 8
35 42
13 21
Boettcherisca peregrina
1
DNA DNA
DNA C ICytochrome c oxidase subunit I
COI
62 100 COI PCR
263 750 bp BioEdit 7.0 MEGA 4
Kimura’s
2-parameter method Neighbor-joining method
COI
A-7
1
DNA D-loop
DNA D-loopD-loopD-loop 5 (SNP)D-loop 3(VNTR)
(STR) 10 D-loop 5SNP 3 D-loop 3VNTR 3 6
STR
STR
A-8
DNADNA extractor FM kitDNAAmpFlSTR Cofiler PCR Amplification
KitSTR
DNADNADNA
0.125ng 32 DNA 0.250.5ng 30
STR 9 10
89% 1
5
DNADNA
A-9
The variation in spatially adjacent pollen assemblages from a
potential assault location
Han Lin1* and Lynne Milne2
1 Centre for Forensic Science, University of Western Australia,
Australia 2 School of Earth and Geographical Sciences, University
of Western Australia, Australia
E-mail: *
[email protected]; Tel: +61 432067779
Abstract Dispersed pollen assemblages recovered from soil and dust
samples are characterised by the
pollen types present and their relative occurrence in a 200 or more
grain count. From this, a
palynologist can qualitatively determine if samples are from the
same locality. Unlike some types of
forensic evidence, no two pollen assemblages will be theoretically
identical because of numerous
taphonomic processes. The disadvantage of this qualitative type of
analysis in a forensic case is the
difficulties in convincing lay persons that minor differences in
pollen assemblages from the same
locality is normal. Although statistical analysis can not replace
qualitative assessment in this
discipline, it may support and validate it in courts.
This project focused on determining the degree of dissimilarity
that can occur between pollen
assemblages from the same site and other localities. Three
replicate soil samples were collected
from each of ten quadrants within a small isolated garden
considered a potential assault location.
Pollen analysis of these samples and soil samples from three
different environments was conducted
and the results statistically analysed. Results indicate that the
natural range of dissimilarity between
replicate, comparator and evidentiary pollen assemblages from the
same locality can be delimited.
These findings will potentially assist in the future presentation
and validation of palynological
evidence in courts.
A-10
Sample
DNA COI
C. megacephalaC.rufifaciesC. pinguisL. porphyrinaO. spinigeraS.
ruficornisS. javanica
(2006/7)
(2005/3-4)
2007/9-10
2007/9-10
21
A-11
)
60-90
(RAPD, Random Amplified Polymorphic DNA)(ITS)5.8S18S26S
RNADNARAPDDNA
(26S ribosome RNA)
PA-12